Professional Documents
Culture Documents
Structure and Function of Laminin: Anatomy Ofa Multidomain Glycoprotein
Structure and Function of Laminin: Anatomy Ofa Multidomain Glycoprotein
Structure and Function of Laminin: Anatomy Ofa Multidomain Glycoprotein
and
function
multidomain
KONRAD
lnstitute
Biocenter
of laminin:
BECK,*
IRENE
HUNTER,T
AND JURGEN
for Biophysics,
University, A-4040 Linz, Austria,
of the University, CH-4056
Basel, Switzerland
is a large
(900
kDa)
mosaic
protein
composed
of many distinct
domains
with different
structures
and
functions.
Globular
and rodlike domains
are arranged
in an extended
four-armed,
cruciform
shape that is
well suited for mediating
between
distant
sites on cells
and other components
of the extracellular
matrix.
The
a-helical
coiled-coil
domain
of the long arm is involved
in the specific assembly
of the three chains (A, Bi, B2,
and possible
variants)
of laminin
and is the only domain composed
of multiple
chains.
It is terminated
by
a large globular
domain
composed
of five homologous
subdomains
formed
by the COOH-terminal
part of
the A chain.
Sites for receptor-mediated
cell attachment and promotion
of neurite outgrowth
reside in the
terminal
region of the long arm. A second cell attachment site, a cell signaling
site with mitogenic
action,
binding
sites for the closely
associated
glycoprotein
nidogen/entactin,
and regions
involved
in calciumdependent
aggregation
are localized
in the short arms.
These
domains,
are composed
of Cys-rich
repeats with limited homology
to EGF, are
the most highly conserved
regions in laminins
of different origin.
At present,
most structural
and functional
tumor,
which
and
dissected
limited
proteolysis.
nins
from
different
for a laminin
expressed
by a
can be readily
isolated
in native
into
functional
fragments
by
Increasing
variations
species
information
on lami-
tissues
demonstrates
Isoforms
of laminm assembled
from different
chains
are focally and
transiently
expressed
and may serve distinct
functions
at early stages of development
even before being laid
down as major components
of basement
membranes.
-BECK,
K.; HUNTER,
I.; ENGEL, J. Structure
and
function
on laminin:
anatomy
of a multidomain
glycoprotein.
FASEB].
4: 148-160;
1990.
considerable
and
of structure.
148
of a
glycoprotein
ABSTRACT
Laminin
anatomy
ENGEL
and tDepartment
of Biophysical
Chemistry,
LAMININS,
A FAMILY
OF LARGE
multidomain
glycoproteins of the extracellular
matrix (ECM),1 have attracted
much interest because of their importance
in the development and maintenance
of cellular organization.
Important cellular functions attributed
to laminin include
stimulation
of growth and differentiation,
neurite
outgrowth promotion,
and mediation
of cell communication. Laminin
is the first ECM protein
detected
during
embryogenesis;
it is present
at the two-cell stage in the
mouse embryo.
In later development
and in mature tissue it serves as a ubiquitous
and major noncollagenous
component
of basement
membranes.
It participates
in
the assembly
of this specialized
form of the ECM and
mediates
cell attachment
and maintenance
of the differentiated
state of epithelial
and endothelial
cell layers
that are intimately
associated
with their
basement
membranes.
There are a number
of excellent
recent
reviews
on
various
aspects
of laminin
and other ECM
proteins
(1-5) and on basement
membranes
(6). In this review
we focus on the structure
of laminin,
the progress
that
has been made in assigning
functions
to distinct
domains of the molecule,
and variations
of the laminin
structure
in tissue-specific
isoforms
and in phylogenetically distant
laminins.
SOURCES
OF
LAMININ
Laminin
was first isolated from the mouse EngelbrethHolm-Swarm
(EHS) tumor (7) and from the extracellular deposit
of murine
parietal
yolk sac (PYS) carcinoma
cells (8). Laminin
from these sources
can be
readily
extracted
and purified
in intact
form under
nonreducing
and nondenaturing
conditions.
In particular, EHS tumor tissue is a rich and convenient
source
of laminin
from which
it is extractable
by neutral
buffers containing
0.5 M NaC1 (9). By a superior
method,
with buffers
nm is extracted
containing
in the form
10 mM EDTA,
Abbreviations:
ECM,
extracellular
matrix;
EGF,
growth
factor;
EHS,
Engelbreth-Holm-Swarm;
HSPG,
sulfate
proteoglycan;
PYS, parietal
yolk sac; SDS-PAGE,
dodecyl
sulfate-polyacrylamide
gel electrophoresis.
0892-663819010004-0148/$01
lami-
with nido-
.50.
epidermal
heparan
sodium
FASEB
gen/entactin
(10). Consequently
most of the biochemical
and biophysical
work has been performed
with EHSlaminin,
which
thus became
the prototype
laminin.
Isolation
of laminins
from
normal
tissues
is often
difficult.
It has been achieved
in many cases only with
denaturing
and reducing agents (5, 11), but extraction
with EDTA appears
to be advantageous
(12). Nevertheless a few tissueand cell-specific
laminins
with distinctly different polypeptide
chains, chain compositions,
and structures
are known today. Although
data on these
so-called isoforms
and on laminin
variants
from phylogenetically
distant
species
are still fragmentary
(see
below),
there
is increasing
evidence
that the wellknown laminin
from EHS tumor is just one member
of
a protein
family.
The reader
should
remember
that
considerable
and
those
variations
that
have
studied
GROSS
STRUCTURE
AND
EHS TUMOR
LAMININ
this laminin
in less detail.
SHAPE
OF
MOUSE
Rotary
shadowing
electron
microscopy
of laminin
revealed an unusual
cruciform
structure
(Fig. l#{192}),
with
three apparently
identical
short arms of 36 6 nm and
a long arm of 77 nm (13). Recent
studies
(14) have
shown, however, that one of the short arms is considerably longer (48 4 nm) than the other two (34 4
nm). The two smaller arms each contain
a central and
a terminal
globule separated
by rodlike regions, whereas
the longer arm contains
an additional
globular
region
(Fig. IA, Fig. 1G, arrows). The long arm of laminin
appears as a rather flexible rod with a large terminal
globule, which at the low resolution
of rotary shadowing
can
be resolved
into two closely spaced
smaller
globules.
Negative
staining
reveals that the globule
adjacent
to
the 3-nm thick rod is composed
of three, and the more
distant globule of two subdomains,
each 4 nm in diameter (Fig. 1B).
Electron
microscopy
thus revealed
a very extended
protein with a maximum
dimension
of 120 nm, consisting of globular
and rocilike elements.
Hydrodynamic
measurements
showed
that this shape
is essentially
preserved
in solution
(13).
FRAGMENTS
OF
LAMININ
Fragmentation
by limited
proteolysis
has been
instrumental
in the elucidation
of the domain
organization and detailed
structure
of large multidomain
proteins such as laminin
and fibronectin.
Laminin
has
been cleaved into a number
of distinct fragments
using
a variety
of enzymes.
Localizations
were derived
by
pro(16,
17). Biochemical
and immunological
studies have enabled the assignment
of functions to distinct domains.
Table 1 is an overview of the important properties and
functions of several well-defined
fragments,
and their
LAM IN IN
C1-4
#{149}.
..
E4
P1
..,
I-*1
.
Figure
1. Electron
micrographs
of EHS-laminin
(A, B) and defined
laminin
fragments
(C-I) after rotary shadowing
(A, G-I) and negative staining
(B-F). Fragments
E3, T8, E8, and C8-9 originate
from the long arm of laminin.
Note that the second terminal
globule at the tip of the long arm is visible in intact laminin
only (double
arrow in A and B) but is absent
in fragments
T8, E8, and C8-9
(D-f). Fragment
C1-4 comprises
the entire short-arm
structures
of
laminin
and fragment
P1 the inner rodlike regions of the three short
arms. Note that one of the short arms is longer than the two others
and contains
a third globular
unit (arrow
in A and C). For further
information
molecule,
on fragments
see Table I and
and their
localization
Fig. 2. (bar: 50 nm).
in the
laminin
localization
within
the laminin
molecule
is shown in
Fig. 2.
The short arms of laminmn are remarkably
resistant
to proteases,
and several fragments
comprising
intact
or truncated
short arms have been detected
(14, 15).
Fragment
P1 is most resistant
to proteases
and comprises the inner region of the cross (Fig. iN). In fragment El, the outer part of one short arm has been removed in the form of fragment
E4 (Fig. 11) and the
other
Fragment
E3 (Fig. 1C), comprising
the outer of the
two globules
at the end of the long arm (G4-G5
in
Fig. 2), is readily released during proteolysis
and is absent from fragments
derived
from the long arm. Fragments T8 (Fig. 1D) and E8 (Fig. 1E) of the long arm
consist of rodlike regions
of different
lengths
together
with the adjacent
globular
domain
(G1-G3
in Fig. 2),
149
TABLE
of mouse EHS-laminin
M (kDa)6
SDS-PAGE
Fragments
native
red.
red.
NH2 terminal
aa
Shape
Short-arm
P1
290
280
El
450
400
E4
75
75
550
550
C1-4
nd
50
other
Predominant
Prominent
60
nd
Aperiodic
Mitogenic
Cell attachment
Binding of nidogen/entactin
Aperiodic
Mitogenic
18
If
Aperiodic
Inhibition
of Ca2-induced
aggregation
of larninin
21
Fig. IC
Aperiodic
Ca2-dependent
14
Fig.
IH
As C1-4,
but with one
truncated
arm
7 (B!)
a
Fig.
nd
Reference
structures
bands
functions
18
18, 19
10, 20
aggregation
Long arm
C8-9
340
E8
190 (A)
+170(BI-B2)
240
+
T8
nd
25K
50
140 (A)
80 (B1-B2)
55
cs-helical
(75%)
Cell attachment
(P. End, M. Bruch,
unpublished
results)
1540 (BI)
1329 (B2)
1886 (A)
Fig. IE
cr-helical
(45%)
Cell attachment
Promotion
of neurite
Outgrowth
nd
1679 (BI)
1473 (B2)
2009 (A)
Fig. ID
12 (B2)
10 (B!)
1679 (BI)
1473 (B2)
50
140 (A)
45 (BI)
+ 35 (B2)
+
80 (A)
25 (Bl-B2)
26 (B1-B2)
+
E3, C3
Fig. IF
190 (A)
+ll0(B1)
+ 90 (B2)
50
nd
Short
rod
Fig. 1C
cr-helical
(30%)
Cell attachment
cs-helical
(100%)
Antibodies
against 25K
inhibit neurite outgrowth
il-structure
19, 22
22, 23
22
15, 20
G1-G3.
Laminin
is unevenly
glycosylated,
average
14 (24,
25) to 25% (26), and therefore
the molar masses of the
native fragments
deviate to different
extents from those
that may be predicted
from the sequence.
Glycosylation
also causes anomalous
migration
of the chains during
SDS-PAGE
giving apparent
molar masses.
Circular
dichroism
of intact laminin
revealed
about
25% a-helix,
which melted
out in a sharp transition
centered
on 59#{176}C
(15, 27). The conformation
of the
short-arm
fragments
cannot
be classified
in terms of
well-known
secondary
structure;
fragments
derived
from the long arm are highly a-helical
(Table 1) and
fragments
E3 and C3 have a 3-structure
(14, 15).
150
Vol. 4
Feb. 1990
A MODEL
OF
MOUSE
EHS
LAMININ
Mouse
EHS tumor laminin
consists of three different
polypeptide
chains-A
(440 kDa),
Bi, and B2 (each
about 220 kDa) -which
are disulfide linked to form the
characteristic
asymmetric
cross-structure
seen by electron microscopy
(Fig. IA). The recent cloning
and sequencing
of all three chains of mouse laminin
(17) as
well as chains from human
(28-30),
Drosophila (31, 32),
and rat s-laminin,
a tissue-specific
variant (11), have revealed a domain
organization
that fits well with the observed ultrastructure,
and have suggested
the arrangement of the three chains within the laminin
molecule
shown in Fig. 2.
All three chains contain
six domains,
1-VI (Fig. 2).
Additional
globular
and rodlike domains
(lila, IVa) are
found in the short-arm
region of the A chain. Domains
I and II, located at the COOH-terminal
ends of the B
chains and in a related
region of the A chain, contain
a series of heptad repeats (33) and are predicated
to be
a-helical.
The calculated
length of these domains,
assuming
a distance
of 0.15 nm/residue
in an a-helix, is
85 nm, which
is in agreement
of the
BECK ET AL.
(A)
ifib
1-4
4
(Th
NH2 (Bi)
VI
ifi
U
In addition
to the six common
domains,
the Bi chain
contains
an additional
40-amino-acid-long
sequence
(domain
a) located between
domains
I and II. It contains 8 Gly and 6 Cys, and is likely to be highly folded
and stabilized
by disulfide
bonds.
It is incompatible
with a-helix
formation
and is presumed
to be looped
out of the long arm, although
no morphological
structure corresponding
to this domain has been detected
by
electron
microscopy.
Domain
G forms
the COOHterminal
region of the A chain with no counterpart
in
the Bi and B2 chains.
Its predicted
structure
fits well
with the large terminal
globule seen by electron
microscopy (Fig. IA). Domain
G consists of five homologous
repeats
(Gi-G5),
which
probably
represent
the five
subdomains
detected
by negative
staining
of intact
laminin
(Fig.
SEQUENCES
Short-arm
Gi
G2
G3
8-9
G4
LJ
3
Figure
2. Structural
model of laminin.
Designations
of domains
by
roman numerals
is according
to ref 17. Cys-rich
rod domains
in the
short arms are designated
by symbols
S and the triple coiled-coil
region (domain
I-LI) of the long arm by parallel
straight
lines. In
the Bl-chain,
the cr-helical
coiled-coil
domains
are interrupted
by a
small Cys-rich
domain
a. Interchain
disulfide
bridges are indicated
by electron
microscopy
(13).
LAMI N IN
1B).
AND
CONFORMATION
structures
To examine
in more detail the relationship
between
the
A, BI, and B2 chains,
the amino acid sequences
of all
three chains were compared.
Alignment
of the shortarm sequences
is shown schematically
in Fig. 3.
The sequence
of the NH2-terminal
domain
VI is
significantly
conserved
in all three chains of EHS laminm, and highly homologous
regions have been found at
the NH2 termini
of all other laminins
sequenced
so far.
In particular,
the 6 or 8 Cys residues
are conserved;
a
sequence
WWQS
in the middle of the region and a pattern of Y(Y/F)YX8G
terminating
region VI are highly
preserved.
The latter motif is also found in a hypervariable region of some immunoglobulins.
The inner globular
regions (domain
IV) of the A and
B2 chains are homologous
and may be considered
to be
derived
from the Cys-rich,
so-called
EGF-like
repeats,
by a large insertion
between
the third and fourth Cys
of one of these motifs (34). Within
the A chain,
this
region occurs twice (IVa and IVb), corresponding
to
the two inner globules
seen on one short arm by electron microscopy
(Fig. IA, Fig. 1G). The large size of the
insertions
(180-200
amino acids, lacking Cys) explains
their appearance
as distinct domains
in electron
micrographs.
Domain
IV of the BI chain has a unique
sequence motif unrelated
to domain
IV of the B2 and A
chains. In the homology
scheme shown in Fig. 3, it appears
as an insertion
(IV) together
with the adjacent
Cys-rich
repeats.
There
are significant
homologies
between
the Cysrich, EGF-like
domains
III and V of all three chains,
and in a single chain
there are homologies
both within
and between
these domains.
The EGF-like
repeats
may
be classified
on the basis of the number
of residues
be-
but
responding
positions
species
well
are considered.
conserved
when
sequences
is clearly
in cor-
from different
demonstrated
by
RYVVLPRP
PCHOC
and domain
IV of the B2 chain. The
partial sequence
of the core protein
of basement
membrane heparan
sulfate proteoglycan
(HSPG)
also contains tandem
repeats with 8 Cys (35) similar to region
IV and the neighboring
repeats of the A and B2 chains
of laminin
(Fig. 3).
Assuming
that the EGF-like
repeats
are arranged
linearly,
an average
translation
per domain
of 2.5 nm
follows from the number
of repeats
and the electron
microscopically
derived
dimensions
of the short-arm
rods.
This
an individual
mation
III1)IIlIIIECPAc
B2
,x
Long-arm
VVV
VV
IU i i i i
i i i
i i i#{149}
1U)
EJSCDDDC
{ ocoDI
LRtiDN
HSPG
3. Alignments
of sequence
regions
in the short-arm
structures of EHS-laminin.
Regions
with predicted
globular
conformation are indicated
by circles
and Cys-rich
EGF-like
repeats
by
squares.
For best alignment
of Cys-rich
repeats,
an insertion
in the
Figure
B! chain (dashed box) and a repeat and gap in the A chain were
introduced.
Cys-rich repeats with additional
insertions (see text)
are indicated by squares with half circles. Domains VI in all three
chains are related (hatched),
whereas
domain
IV (cross-hatched)
in
chain BI is unrelated to domains IV in A and B2. Cys-rich repeats
of heparan sulfate proteoglycan
(HSPG, 35) are aligned for comparison. Sequence data for mouse laminin are by Sasaki et al. (17).
Triangles mark putative sites of N-linked
glycosylation.
Sequence
regions for which functions have been proposed (Table 2) are indicated by their sequences in one letter code. Sequences that are probably involved in disulfide linkage of the three chains (Fig. 2) are indicated at the right-hand
side.
(34).
structure
(33).
Using
the long-arm
152
Vol. 4
of the
Feb. 1990
BI
chains
fragments
C8-9
and
E8 (Fig.
2),
we have recently
shown
that the a-helical
domains
I and II are involved in chain assembly (36). When the
A, Bi, and B2 chains present
in these fragments
are
separated
and unfolded
in urea, they reassemble
into
molecules
which
in their a-helix
content,
apparent
molar mass, chain composition,
and ultrastructural
appearance
are indistinguishable
from the native
fragments. The results indicate
that all three chains interact
to form a triple-coiled
coil, which can extend the length
of the long arm. The highly specific nature of this interaction suggests that it is the mechanism
by which laminm assembles
in vivo.
Figure
4 shows
a cross-section
through
a triple
coiled-coil,
as is suggested for laminin.
Burying the
hydrophobic
residues located in positions a and d in the
center of the structure,
thus shielding them from the
aqueous environment,
is energetically
favorable and is
the driving force for coiled-coil
formation.
Hydrophiic
amino acids are exposed on the surface, and the coiled-
coil is further
of mouse
EHS tumor
laminin,
Drosophila laminin,
and s-laminin
(11, 31). This
observation
suggests
that the order
of EGF-like
domains is of functional
importance
and argues against a
simple
structural
role as spacer elements.
The functional significance
of the short-arm
structures
is further
emphasized
by their high degree of conservation
(about
60% on average)
in laminins
that exhibit only 20% homology
in regions
of the long arm (ii, 31).
Within mouse EHS laminin,
the two EGF-like
repeats
adjacent
to each side of domain
IVa of the A chain are
similar
to those in the same position
around
domain
a comparison
In contrast
to the short arms, the sequences
of the A,
Bi, and B2 chains assigned
to the rodlike region of the
long arm (domains
I and II) have little sequence
homology,
but all contain
a heptad
repeat
of the type
(a,b,c,d,e,f,g)n
where hydrophobic
amino acids are located preferentially
in positions
a and d, charged
residues normally
in positions
e and g, and polar residues
frequently
in b, c, and f. Such sequence
motifs are
characteristic
of proteins
in which
two or three
ahelices are wound around
each other to form a coiledcoil
agrees
well
domain,
residues
e and
stabilized
by ionic interactions
between
g.
BECK E AL.
[nm]
0.5
looped
helical
domains
matched,
all
in parallel
of
(Fig.
5), which
Cys-residues
is in agreement
aligned
with
their
based
on the preference
cific positions
which
F,
Figure
4. Projection
of a three-stranded
a-helical
coiled-coil,
one
heptad
each, viewed from the NH2 terminus.
A distance
of 1.0 nm
is assumed
between
the helix axes. Positions
of 3-carbon
atoms
within
a heptad
repeat
assuming
3.6 residues
per turn (a-g).
The
clustering
of hydrophobic
side chains (filled circles) and electrostatic
interactions
(dashed
circles) are schematically
indicated.
Side chain
in positions
b, c, and f are located
at the surface
and are normally
polar or charged
(open circles).
within
of certain
the heptad
is a measure
amino
acids
for spe-
of the probability
of forming
coiled-coil structure.
The lowest probability
for coiledcoil is predicted
for all three chains near the NH2terminus of domain II, and this region is particularly
susceptible
to proteolysis
(14). The highest value of F#{176}
is found for the Bi chain near its COOH-terminus.
The entire B2 chain exhibits intermediate
values, and
the lowest values are found for the A chain. Thus,
although all three chains of laminin are believed to par-
V25K
B2dJ
Bi:
LQQSAA
KVESLI
B2:
NDILNN
NSVSSL
AHVHSN
PHQ
B2
EPA
SDI
LHREHG
Bi
Bi
uE8
:i
[j
100
200
300
Figure
5.
Putative
factor
arrangement
500
400
I
ii:
coiled-coil
SRARK
V//////DO.5
F1 x 108:
of the A, Bi, and
B2 chains
within
i..i>O2
the long
arm
I
of EHS
laminin.
1<0.2
Cys
pairs
at the
NH2
lOnm
terminus
of
domain
II are probably
involved
in interchain
disulfide
bonding
(see also Fig. 3). The established
disulfide
bridge near the COOH
termini
of the Bl and B2 chains is indicated
by a vertical
bar. Domain
a is looped
out. As originally
noted by Barlow
et al. (37), several phase
shifts have to be introduced
in the heptad repeats and these are indicated by displacements
between
blocks.
Proline
residues
that tend
to disrupt a coiled-coil structure are indicated by filled circles. Blocks are marked according to their coiled-coil potential as expressed by
the probability
factor F defined by Parry (38). Putative sites for N-glycosylation
are indicated by triangles.
For some sites (filled triangles), evidence for glycosylation was obtained (39). Sequence regions claimed
to be involved
in neurite
outgrowth
activity
are indicated
by horizontal bars and their first five residues (Table 2). The number of residues in the coiled-coil region is indicated (abscissa). NH2terminal sequences of fragments E8, 25K, and E3 are indicated. The length of coiled-coil regions and diameters
of domain
G are drawn
approximately
to scale.
LAMININ
153
ticipate
in coiled-coil
formation
(see above),
in homotypic interactions
the A chain would not be expected
to
adopt
this structure.
This is in agreement
with recent
observations
from this laboratory
(I. Hunter
andJ.
Engel, unpublished
results) that although
the Bi and B2
chains alone can form a coiled-coil,
the A chain can
participate
in coiled-coil
formation
only in the presence
of the B chains.
Domains
I and II of all three chains together
account
for a protein
molar mass of 192.8 kDa. Assuming
a
length of 77 nm for the long arm, a mass-per-length
ratio of 2500 Da/nm
follows, which is slightly higher
than expected
for a perfectly
packed
triple-coiled-coil
(2250 Da/nm)
and suggests
some nonhelical
interruptions, presumably
due to the variations
in the stability
of the coiled-coil
regions
described
above, and to the
presence
of a number
of proline
residues
(Fig. 5).
The COOH-terminal
region of the A chain adjacent
to the coiled-coil
domain
contains
five internal
homologous repeats
(G1 to G5), each consisting
of about 200
residues.
For human
laminin,
only regions
G2 to G5
have been sequenced
(30) and exhibit an overall identity of 74% with mouse EHS laminin.
This homology
is surprisingly
low when compared
with more than
90% identity
of other parts of mouse and human
laminm (28, 29).
readily
extractable
EDTA-containing
from
buffers
basement
short arms
21, 50, 51).
at the level
laminin
is
membranes
with
(10).
Laminin
forms a particularly
stable complex with
nidogen/entactin,
which can be extracted from mouse
EHS tumor
as a 1:1 complex
(10). Nidogen
has been
observed
to bind to the Cys-rich,
EGF-like
repeats
(domain
III) (10, 20) of both the B! and B2 chains
responsible
electron
short
for binding
microscopy
(domain
VI)
have been
to the terminal
and
long
localized
globules
(domain
G1-G5)
by
of the
arms
(50).
GLYCOSYLATION
Seventy-four
A major binding
potential
N-glycosylation
in the three
sites
chains
NXS/T
stability
against
proteases
STRUCTURE-FUNCTION
(42).
RELATIONSHIPS
Laminin
exhibits
a variety
of biological
activities,
including
promotion
of cell attachment,
growth
and
differentiation
of a number
of cell types, and multiple
interactions
with other basement
membrane
components (1-6). Progress
toward assigning
these functions
antibodies.
fragments
Table
1 provides
a summary
the localization
of these fragments
within
molecule
is shown in Fig. 2. In addition,
154
Vol. 4
Feb. 1990
of the
and
the laminin
the recent
terminal
globule
arm
was localized
(15) and
may
to the
also be
membrane
creating
components,
and
maintaining
of laminin
is ideally suited
of a variety
of basement
complex
3-dimensional
surface
of effects
the basement
receptors
through
on cellular
membrane
which
and
contact
cell-
processes.
good
tions
agreement
with immunoultrastructural
(58) indicating
that the inner regions
BECK ET AL.
TABLE
Chain
Domain
Bi
aa-residues
Peptide
442-447
(H:
5:
D:
B!
IV
64 1-660
Proposed
III
902-906
B!
III
925-933
function
Reference
LGTIPG
LGTIPG
RGTVPG
LGTLNN)
RYVVLPRPVCFEKGMNYTVR
Binding
RYVVLPRPVCFEKGTNYTVR
5: RVLVFPRPVCLEPGLSYKLK
D: RQVVALNEVCLEAGKVYKFR)
B!
chains
of heparin;
promotes
cell adhesion
(H:
of murine melanoma
cells, fibrosarcoma,
glioma, pheochromocytoma,
and aortic
endothelial
cells
PDSGR
(H: PDSGR
s: PGSQR
D: VASGL)
Similar to (CDPG)YIGSR
(see below);
proposed to act synergistically
with
YIGSR
CDPGYIGSR
CDPGYIGSR
5: CRAGYTGLR
D: CQEGYSGSR)
(H:
Promotion
of cell attachment,
chemotaxis,
neuronal attachment
but not neurite
outgrowth
44, 45
Promotes melanoma
cell migration
Inhibits formation of lung tumors
44, 46
44, 47
1542-155!
IlIb
1118-1128
GTFALRGDNGQ
2091-2 108
SRARKQAASIKVAVSADR
For
sequence
43
44
B2
Drosophila)
RNIAEIIKDA
(H: RDIEEIMKDI
D: RELKDEVQ.NI)
58a
Stimulates
neurite
outgrowth
48
cells; can
49
49
localization,
see Fig. 3 and Fig. 5.
bThe homologous
sequences at corresponding
sites of other laminins (H, human;
s, s-laminin;
D,
that have not been tested for their activity are indicated in parentheses.
The sequence of the synthetic peptide corresponds
to the partial
reported in (37); in the complete sequence (17), N is replaced by D.
interior
of basement
epitopes
are located
basement
membrane
surface.
Laminin is a potent stimulator
memat the
A general problem
associated
with studies using
thetic peptides
is that they represent
only a small
of the domains
of neurite
outgrowth
likely
therefore
to have
the correct
structure.
Many
the functions
mapped
shown
to be conformation
rounded by laminin-producing
Schwann cells. Domainspecific antibodies
have implicated
sequences
within
fragment E8 as important
for this activity, and a synthetic peptide corresponding
to a defined sequence of
the B2 chain (Table 2) has been reported to have neu-
mitogenic
function
of fragment
P1 is lost upon reduction (P. End, unpublished
results),
and cell adhesion
rite
to have
sequences
must also consider
the structural
requirements of such sequences.
To achieve this, studies (44)
were performed
recently
with a cyclized
form of the
peptide YIGSR.
Homology searches have not been helpful in suggesting other possible functions of distinct regions of lami-
dence is comparable
to that of EGF (18). This function
has been localized
to fragment
P1, which consists
of
Cys-rich
repeats
(domain
III) and is not related
to its
outgrowth-promoting
cell attachment
(Table
activity
(48).
properties
(18). Synthetic
Another
pep-
rodlike
shown
region
peptides
cor-
neurite
abolished
outgrowth
upon
ture studies
specific
domains
have
dependent.
In particular,
of
and
to
synpart
activities
denaturation
aimed
been
the
of fragment
E8 are
57). Therefore,
fudelineating
functional
(20,
at precisely
responding
to domain
III sequences,
including
the pentapeptide
YIGSR,
which represents
part of the Cysrich repeat with closest homology
to EGF, had no mito-
mentioned.
Weak homologies
are also observed between domains I and II and many other proteins
con-
genic activity
homology
LAMININ
(18).
taining
coiled-coil
exists
structures.
between
Perhaps
domains
GI
an
interesting
to G5
and
sex
155
steroid-binding
protein
(codes G08607
and S00077 of
the National
Biomedical
Research
Foundation
protein
sequence
data base).
In agreement
with its numerous
biological
activities,
a variety of cell-surface
receptors
necessary
to mediate
laminin
activity
have been identified
(2, 59). In the
present context the fast-growing
field of laminin
receptors cannot be reviewed,
and only a few examples
will
be mentioned.
A 67-kDa protein is particularly
well expressed
in metastatic
cells (60), which could be eluted
from a laminin
affinity
column
by the domain
IIIderived,
synthetic
peptide
YIGSR
(45). Recent studies
have questioned
this proposed
binding
site, as purified
67-kDa
receptor
tron
microscopy
was
shown
to bind
by rotary
specifically
shadowing
to the
elec-
top of the
AND
STRUCTURAL
VARIANTS
LAMININ
The identification
of distinct
but related
pepsin fragments in tissue digests as compared
with digests of EHS
laminin
(64), antigenic
differences
between
laminins
derived
from tumor
cells and normal
tissues (12, 65),
and the absence or reduced
expression
of the 440-kDa
A chain in a number
of cell types in culture (5, 6, 30),
has led to the suggestion
that structural
variants
of
laminin
may exist. In addition,
variation
in the levels
of mRNA
for the A, BI, and B2 chains of laminin
in
different
tissues of the same species (66, 67) indicates
the formation
of tissue-specific
isoforms,
possibly with
different
functional
properties.
Laminin
appears
to be developmentally
regulated.
During
mouse
embryogenesis,
the B chains
are detected at the 2-4 cell stage, whereas
the 440-kDa
A
chain is not detected
until the 16-cell stage (68), which
156
Vol. 4
Feb. 1990
is coincident
with the ultrastructural
appearance
of a
distinct
basement
membrane.
During
normal
kidney
development,
antibodies
against
EHS laminin
could
detect only B chains in undifferentiated
mesenchyme,
but on conversion
to a polarized
epithelium,
A chain
epitopes
were expressed
(69). Whether
these laminin
variants
consist
of B chains
only or have in addition
a
modified
A chain,
antigenically
unrelated
to the A
chain
of EHS
laminin,
is presently
unknown.
Isolation
and
characterization
of laminin
from
Schwannoma
cells, which lack the 440-kDa
A chain,
revealed
a Y-shaped
molecule
with one long and two
short arms (Table 3). Although
only B chains (200-220
kDa) could be detected
by SDS-PAGE
under reducing
conditions,
the Schwannoma
laminin
had an apparent
molecular
mass of about 850 kDa under nonreducing
conditions,
indicating
that it was not formed
solely
from the Bi and B2 chains and suggesting
the existence
of a variant (shorter)
A chain. Such variant
forms have
recently
been described
in laminin
from mouse heart
(300 kDa) (12) and 3T3-Li
cells (180 kDa) (75). In
3T3-L1 cells, as shown previously
for cells synthesizing
the 440-kDa
A chain, B1-B2 disulfide-linked
dimers are
formed as intermediates
in laminin
assembly,
but these
can be secreted
only in the form of a ternary
complex
with the 180-kDa
A chain (75). Merosin
first described
as an 80-kDa
protein
present
in basement
membranes
of human
muscle
and peripheral
nerves
(76) has been
identified
on the basis
of a 40%
homology
as a fragment
originating
from the COOH-terminal
portion
of
another
tissue-specific
A chain variant (71). This new A
norsimi-
lar
to EHS
laminin
(71).
All isoforms
investigated
so far by electron
microscopy
(12, 22, 70) have a long-arm
terminal
globule,
which
in mouse
EHS laminin
is derived
entirely
from
the A chain.
Available
evidence
structural
variants
of laminin
suggests
therefore
that
are of the type A(B)2.
Whether
the A-chain
variants
detected
so far are
related to the 440-kDa
A chain, possibly splicing variants, or are novel subunits,
awaits the sequencing
of
these chains.
Additional
structural
information
has come from
studies of laminins
of different
species origin. The gross
shape
of laminins
originating
from different
phyla
ranging
from Cnidaria
to mammals
has been investigated in some detail by electron
microscopy
(Table 3).
All of the laminins
studied exhibit an asymmetric
crossstructure
with one long arm bearing
a terminal
globule
and three short arms each with terminal
and centrally
located globules.
The structure
of these molecules
thus
closely resembles
that of mouse EHS tumor
laminin.
The dimensions
of the short arms are well preserved
(Table 3) but the length of the long arm varies considerably
with species. Two globular
domains
at the tip
of the long arm, connected
by a short, 5-10 nm long
rod, may be related
to the subdomains
G1-G3
and
G4-G5
seen in negatively
stained
preparations
of
mouse tumor laminin
(Fig. 1B). The preparation
of a
BECK ET AL.
TABLE
Mouse
Mouse
sources
Shape
EHS tumor
heart
Chains
Prominent
unrelated
but possible
structures
laminin,
Rat Schwannoma
Human
cells
placenta
Y-shaped
Similar
to EHS laminin;
long arm,
83 nm
Sea urchin
embryo
Anthomedusa
Length
reduction.
measurements
to A and
of molecules
globules
only
14
12
22,
70
71
immunologically
B chains
400 kDa
exhibits
capsules
subpopulation
Leech ganglion
13,
B2)
related
Drosophila K, cells
Reference
(A)
72
B2)
73
and
two inner
73, 74
73
were performed
after rotary
shadowing
electron
proteolytic
fragment
from sea urchin laminin,
which is
indistinguishable
by electron
microscopy
from fragment E8 of mouse tumor laminin
(73), indicates
that
there
may
be similarities
between
these
COOHterminal
regions in both laminins.
Thus, modifications
leading
to the elongation
of the long arm in many species may be restricted
to the proximal
part of this arm.
Such length variations
may be required
to span basement membranes
of different
thicknesses.
OUTLOOK
microscopy.
Apparent
molar
masses
were determined
by SDS-PAGE
after
A key problem
for future research
on laminin
will be
the further
elucidation
of structure/function
relationships.
Assignment
of functions
to regions
in the
molecule
by the proteolytic
fragmentation
approach
has been very successful
and will probably
continue
to
be so. Its limitations
are that defined fragments
cannot
be prepared
from all parts of laminin
and the fact that
small fragments
do not retain their native conformation. The alternative
approach
of testing the activity of
peptides
synthesized
according
to sequence
regions suspected to be involved in functions
has become popular
1.AMININ
native conformation
and for functionality.
It is therefore required
to confirm the results obtained
with peptides by independent
methods.
Site-directed
mutagenesis and functional
tests of the modified
laminins
in vitro
157
efforts should
be made to understand
the functional
variations
of the various isoforms
on a structural
basis.
In addition,
there are many
domains
in laminin
to
which
functions
have been assigned
on the basis of
weak arguments
or for which no functions
have yet
been found. Many exciting
discoveries
are expected.
We thank
Ms.
C. Fauser
for
expert
technical
assistance
chain
has a unique
basement
membrane
J. Biol. C/tern. 263,
18. Panayotou,
G., End, P., Aumailley,
M., Timpl,
R.,
gel,
(1989) Domains
of laminin
with growth-factor
Cell 56, 93-101
19. Aumailley,
M., Nurcombe,
V., Edgar,
D., Paulsson,
J.
(Be 1143/1-1).
20. Timpl,
Edgar,
Methods Enzymol.
G. R., Timpl,
proteins:
R., and
molecular
Kuhn,
R. A. (1986) Structures
M. (1986) Structure,
of basement
27.
Robey,
G. R.
P., Rennard,
S. I.,
(1979) Laminin-a
j Biol. C/tern. 254,
28.
Vergnes,J.
P., Braginof a basement
memin culture
by a mouse
and Fujiwara,
S. (1987)
Basement membranes.
Methods Enzymol. 145, 363-391
10. Paulsson,
M., Aumailley,
M., Deutzmann,
R., Timpl,
R.,
Beck, K., and Engel, J. (1987) Laminin-nidogen
complex.
Extraction
with chelating
agents
and structural
characterization.
Biochern. 166,
11-19
11. Hunter,
D. D., Shah, V., Merlie,
J. P., and Sanes, J. R. (1989)
A laminin-like
adhesive
protein
concentrated
in the synaptic
cleft of the neuromuscular
junction.
Nature (London) 338,
229-234
12. Paulsson,
M., and Saladin,
K. (In press) Mouse
heart laminin:
purification
of the nature
protein
and structural
comparison
14.
15.
16.
17.
158
fibronectin,
two multifunctional
matrix. J. Mol. Biol. 150, 97-120
Bruch, M., Landwehr,
R., and Engel, J. (1989) Dissection
of
laminin
by cathepsin
G into its long arm and short arm structures and localization
of regions
involved
in calcium
dependent
stabilization
and self-association.
Eur j Biochern. 185, 271-279
Ott, U., Odermatt,
E., Engel, J., Furthmayr,
H., and Timpi,
R.
(1982) Protease
resistance
and conformation
of laminin.
Eur. j
Biochern. 123, 63-72
Hartl, L., Oberb#{228}umer,I., and Deutzmann,
R. (1988) The N
terminus
of laminin
A chain is homologous
to the B chains.
Eur.
J. Biochern. 173, 629-635
Sasaki,
M., Kleinman,
H. K., Huber,
H., Deutzmann,
R.,
Yamada, Y. (1988) Laminin,
a multidomain
protein.
The A
Vol. 4
Feb. 1990
and
26.
development,
mt. Rev. Exp.
membranes.
glycoprotein
from basement
membranes.
9933-9937
8. Chung,
A. E.,Jaffe,
R., Freeman,
I. L.,
ski, J. E., and Carlin,
B. (1979) Properties
brane
related glycoprotein
synthesized
embryonal
carcinoma-derived
cell line.
9. Timpl,
R., Paulsson,
M., Dziadek,
M.,
25.
K. (1988) Basement
and function.
Adv. Pro-
structure
J.
complex.
Ann. NY
23. Edgar,
D., Timpi,
R., and Thoenen,
H. (1984) The heparinbinding
domain
of laminin
is responsible
for its effects on neurite outgrowth
and neuronal
survival.
EMBOJ.
3, 1463-1468
24. Arumugham,
R. G., Hsieh, T. C. -Y., Tanzer, M. L., and Lame,
145, 3-78
R. (1989) Structure
and biological
activity
of basement
membrane
proteins.
Eur. j Biochern. 180, 487-502
3. Martin,
G. R., and Timpi,
R. (1987) Laminin
and other basement membrane
components.
Annu. Rev. Cell Biol. 3, 57-85
4. Paulsson,
M. (1987)
Noncollagenous
proteins
of basement
membranes.
Collagen Relat. Res. 7, 443-461
Eur.
and
fraghigh
R., Aumai!ley,
M., Gerl, M., Mann,
K., Nurcombe,
V.,
D., and Deutzmann,
R. (1989) Structure and function of
the laminin-nidogen
2. Timpl,
and molecular
M.,
106, 1299-1306
membrane
and Enactivity.
21. Schittny,
J. C., and Yurchenco, P. D. (In press) Terminal short
arm domains
in the basement
membrane
protein
laminin
are
critical
for its self-assembly.
j Cell Biol.
22. Edgar,
D., Timpl,
R., and Thoenen,
H. (1988) Structural
requirements
for the stimulation
of neurite
outgrowth
by two variants of laminin
and their inhibition
by antibodies.
j Cell Biol.
REFERENCES
5. Martin,
with the
B chains.
and
M. Bruch
and M. Paulsson
for valuable
discussions
and communicating
unpublished
results. Original
work from this laboratory
reported
herein has been supported
by The Swiss National
Science
Foundation
(grant
no. 31-9088.87
to J. E.). The present
work by
K. B. is supported
by the
Deutsche
Forschungsgemeinschaft
globular
domain
and homology
proteoglycan
and the laminin
16536-16544
of othe asparigine-linked
sugar chains
of laminin.
Biochirn. Biophys. Acta 883, 112-126
Fujiwara,
S., Shinkai,
H., Deutzmann,
R., Paulsson,
M., and
Timpl,
R. (1988)
Structure
and
distribution
of N-linked
oligosaccharide
chains
on various
domains
of mouse
tumour
laminin.
Biochem. j 252, 453-461
Knibbs, R. N., Perini, F, and Goldstein,
I. J. (1989) Structure
of the major concanavalin
A reactive oligosaccharides
of the cxtracellular
matrix
component
laminin.
Biochemistry
28,
6379-6392
Paulsson,
M., Deutzmann,
R., Timpl,
R., Dalzoppo,
D., Odermatt, E., and Engel,
(1985) Evidence
for coiled-coil
a-helical
regions
in the long arm of laminin.
EMBOJ
4, 309-316
Pikkarainen,
T, Eddy, R., Fukushima,
Y., Byers, M., Shows,
T., Pihlajaniemi,
T., Saraste, M., and Tryggvason,
K. (1987)
Human
laminin
B1 chain.
A multidomain
protein
with gene
(LAMB1) locus in the q22 region of chromosome
7. j Biol.
J.
C/tern. 262,
10454-10462
29. Pikkarainen,
T., Ka!lunki,
Human
laminin
B2 chain.
T,
and
Tryggvason,
K. (1988)
Comparison
of the complete
amino
acid sequence with the B! chain reveals variability
in sequence
homology
between
different
structural
domains.
j BioL C/tern.
263, 6751-6758
30. Olsen, D., Nagayoshi,
T, Fazio, M., Peltonen, J., Jaakkola,
S.,
Sanborn,
D., Sasaki,
T., Kuivaniemi,
H., Chu, M. -L., Deutzmann,
R., Timpl,
R., and Uitto, J. (1989) Human
laminin:
cloning and sequence
analysis of cDNAs encoding
A, BI and B2
chains,
and expression
of the corresponding
genes in human
skin and cultured
cells. Lab. Invest. 60, 772-782
31. Montell,
D. J., and Goodman,
C. S. (1988) Drosophila substrate
adhesion
molecule:
sequence
of laminin
B1 chain reveals domains of homology with mouse.
Cell 53, 463-473
33.
34.
35.
36.
mouse,
264, 1543-1550
Parry, D. A. D. (1987) Fibrous
protein
structure and sequence
analysis.
In Fibrous Protein Structure (Squire,
J. M., and Vibert,
P. J., eds) pp. 141-17!, Academic,
London
Engel, J. (1989) EGF-like
domains
in extracellular
matrix
proteins: localized
signals
for growth
and differentiation?
FEBS
Let:. 251, 1-7
Noonan,
D. M., Horigan,
E. A., Ledbetter,
S. R., Vogeli, G.,
Sasaki,
M., Yamada,
Y., and Hassell, J. R. (1988) Identification
of cDNA
clones encoding
different
domains
of the basement
membrane
heparan
sulfate
proteoglycan.
J. Biol. C/tern. 263,
16379-16387
Hunter,
I., Schulthess,
T., Bruch,
M., Beck, K., and Engel, J.
(In press) Evidence
for a specific mechanism
of laminin assem-
bly. Eur. j
Biochem.
BECK Er AL.
37. Barlow,
D. P., Green,
N. M., Kurkinen,
M., and Hogan,
B. L. M. (1984) Sequencing
of lam
B chain cDNAs
reveals
C-terminal
regions of coiled-coil
alpha helix. EMBOJ
3, 23552362
38. Parry, D. A. D. (1982) Coiled-coils
in a-helix-containing
proteins: analysis
of the residue
types within the heptad
repeat and
the use of these data in the prediction
of coiled-coils
in other
proteins.
Biosci. Rep. 2, 1017-1024
mm
39. Deutzmann,
40.
41.
42.
43.
44.
R., Huber,
H., Schmetz,
K. A., Oberb#{228}umer,I.,
and Hartl,
L. (1988) Structural
study of long arm fragments
of
laminin.
Evidence
for repetitive
C-terminal
sequences
in the Achain, not present
in the B-chains.
Eur. j Bloc/tern. 177, 35-45
Dennis,
J. W., Waller, C. A., and Schirrmacher,
V. (1984)
Identification
of asparagine-linked
oligosaccharides
involved
in
tumor
cell adhesion
to laminin
and type IV collagen.
j Cell
Biol. 99, 1416-1423
Wu, C., Friedman,
R., and Chung,
A. E. (1988) Analysis
of the
assembly
of laminin
and the laminin
entactin
complex
with
laminin
chain
specific
monoclonal
and polyclonal
antibodies.
Biochemistry
27, 8780-8787
Howe,
C. C. (1984) Functional
role of laminin
carbohydrate.
Mol. Cell. Biol. 4, 1-7
Charonis,
A. S., Skubitz,
A. P. N., Koliakos,
G. G., Reger,
L. A., Dege, J., Vogel, A. M., Wohihueter,
R., and Furcht,
L. T. (1988) A novel synthetic
peptide
from the BI chain
of
laminin
with heparin-binding
and cell adhesion-promoting
activities. j Cell Biol. 107, 1253-1260
Kleinman,
H. K., Graf,J., Iwamoto, Y., Sasaki, M., Schasteen,
C. S., Yamada,
Y., Martin,
F A. (1989)
Identification
of a second
active site in laminin
for promotion
of cell adhesion
and migration
and inhibition
of in vivo melanoma lung colonization.
Arch. Biochern. Biophys. 272, 39-45
46.
47.
48.
49.
50.
51.
52.
53.
54.
55.
Y., Sasaki,
M., Martin,
G. R., Kleinman,
LAMININ
Dale, M. (1985)
27, 317-325
56. Nurcombe,
Biological
activities
V., Aumailley,
J. Cell Bloc/tern.
of laminin.
M., Timpl,
D. (1989)
The high-affinity
binding
of laminin
to cells. Assignation
of a
major cell-binding
site to the long arm of laminin
and of a latent
cell-binding
site to its short arms. Eur j Bioc/tern. 180, 9-14
57. Goodman,
S. L., Deutzmann,
R., and von der Mark, K. (1987)
Two distinct cell-binding domains in laminin can independently
promote nonneuronal
cell adhesion and spreading. j Cell BioL
105, 589-598
58. Schittny,
J. C., Timpl, R., and Engel, J. (1988) High resolution
immunoelectron
microscopic
localization
of functional
domains
of laminin,
nidogen,
and
heparan
sulfate
proteoglycan
in
epithelial
basement
membrane
of mouse
cornea
reveals different topological
orientations.
j Cell BIoL 107, 1599-1610
58aMecham,
R. P., Hinek,
A., Griffin,
G. L., Senior,
R. M., and
Liotta,
L. A. (1989) The elastin
receptor
shows structural
and
functional
similarities
to the 67-kDa
tumor
cell laminin
receptor. j BioL C/tern. 264, 16652-16657
59. Edgar,
D. (1989) Neuronal
laminin
receptors.
Trends Neurosci.
12, 248-251
60. von der Mark, K., and K#{252}hl,
U. (1985) Laminin
tor. Bioc/tirn. Biophys. Acta. 823, 47-160
61. Hall, D. E., Frazer,
(1988) Isolation
and
K. A., Hann,
characterization
E. (1988)
Fibronectin
L., Engvall,
E., and Ruoslahti,
E.
receptor
is a member of the integrin
Dillner,
laminin
64. Risteli,
5. (1989)
RuGli
cell line
not of human
R. (1981) Isolation
and characterization
of pepsin fragments
of laminin
from human
placental
basement
membranes.
Bloc/tern. J. 193, 749-755
65. Wewer,
U. M., Tichy,
D., Damjanov,
A., Paulsson,
Damjanov,
ori-
342-343
I. (1987) Distinct
and
M.,
renal
and
antigenic characteristics
of murine
Develop. BioL 121, 397-407
parietal
yolk sac laminin.
66. Boot-Handford,
R. P., Kurkinen,
M., and Prockop,
D. J. (1987)
Steady
state levels of mRNAs
coding
for the type IV collagen
and laminin
polypeptide
chains of basement
membranes
exhibit
marked
tissue-specific
stoichiometric
variations
in the rat.
j Biol. C/tern. 262, 12475-12478
67. Kleinman,
H . K., Ehihara,
I., Killen, P. D., Sasaki,
M., Cannon, F B., Yamada,
Y., and Martin,
0. R. (1987) Genes for
basement
membrane
proteins
are coordinately
expressed
in
differentiating
F9 cells but not in normal
adult murine
tissues.
Dcv. BioL 122, 373-378
68. Cooper,
A. R., and MacQueen,
H. A. (1983) Subunits
of lasninm are differentially
synthesized
on mouse eggs and early embryos. Dev. Biol. 96, 467-471
69. Klein,
G., Langegger,
M., Timpl,
R., and Ekblom,
P. (1988)
M.,
Martinez-Hernandez,
N. A. (1985) Comparative
placental
membrane
with
A., Ohno,
study
laminin
N., and
Kefa!ides,
found
in normal
of laminin
of neoplastic
J. H.
tion. j
73. Beck,
origin.
In Base-
Science
Publ.
L. I., Campbell,
A. G., Duncan,
K. 0., and Fessler,
(1987) Drosophila laminin:
characterization
and localiza-
McCarthy,
R. A., Chiquet,
M.,
Masuda-Nakagawa,
L., and Schlage, W. K. (1989) Structure of the basement membrane protein laminin: variations on a theme. In Cytos/celetal and
Extracellular Proteins: Structure, Interactions and Assembly (Aebi, U.,
and
Engel,
J.,
eds)
pp.
102-105,
Springer-Verlag,
Berlin
74. Chiquet,
tachment
sprouting
M., Masuda-Nakagawa,
to an endogenouis
by leech neurons.
j
L., and
laminin-like
75. Aratani,
Y., and Kitagawa,
Y. (1988) Enhanced
synthesis
and
secretion
of type IV collagen
and entactin
during
adipose
conversion
of 3T3-Lll
cells and production
of unorthodox
laminin
complex.
J. BioL Chern. 263, 16163-16169
160
Vol. 4
Feb. 1990
109, 4a
BECK Er AL.