Cellular Transport

Types of Cellular Transport

Passive Transport
Cell doesn’t use energy
1. Diffusion
2. Facilitated Diffusion
3. Osmosis

Active Transport
Cell does use energy
1. Protein Pumps
2. Endocytosis
3. Exocytosis
 Diffusion & Osmosis Osmosis
Diffusion

Solute moves DOWN the concentration

gradient. Movement of water across a
HIGH to LOW membrane.
Moves from HIGH water
concentration to LOW water
concentration.
 Cells in Solution
Isotonic
A solution whose solute concentration
is the same as the solute
concentration inside the cell.

Hypotonic
A solution whose solute concentration
is lower than the solute concentration
inside a cell

Hypertonic
Hypertonic Normal saline Hypotonic
A solution whose solute concentration (1.1% salt) (0.9% salt) (0.7% sale)
is higher than the solute concentration
inside a cell.
 Types of Transport Across the Membrane

Simple Diffusion Facilitated Diffusion Active Transport

Doesn’t require energy Some carrier proteins do Requires energy or ATP
Moves high to low not extend through the
concentration membrane. Not dependent on
Example: Oxygen or water They bond and drag concentration gradient
diffusing into a cell and molecules through the
carbon dioxide diffusing lipid bilayer and release
out. them on the opposite side.
 Active Transport- Pinocytosis
Cell forms an invagination
Materials dissolve in water to be brought into cell
Called “Cell Drinking”
 Active Transport--Phagocytosis
Cells form phagocytic cup
Used to engulf large particles such as food, bacteria,
etc. into vesicles.
Called “Cell Eating”
 Receptor-Mediated Endocytosis

Some integral proteins have receptors on their surface to

recognize & take in hormones, growth factors, cholesterol, etc.
 TYPES OF TRANSPORT AND
PROCESSING OF RECEPTOR-
LIGAND SYSTEMS

• TYPES : • EXAMPLES:
• Receptor recycled; • Mannose receptor,
ligand to lysosome. LDL receptor.
• Receptor recycled; • IgG & Transferrin
ligand recycled. receptor.
• Receptor demise; • EGF & Insulin
ligand to lysosome. receptor
• Receptor demise; • IgA
ligand not to
lysosome.
Receptor Recycled: Receptor Recycled:
Ligand to Lysosome Ligand also Recycled
Receptor demise: Ligand to Lysosome Transcytosis (IgA)
Steps in receptor mediated
endocytosis
A. Selection of ligand for uptake
I. Receptor tails
II. Adaptors

B. Formation of vesicles
I. Invagination
II. Membrane scission

C. Targetting to endosomes
I. Uncoating
II. Docking and fusion

D. Sorting of ligand & receptors

 Topology of different Receptors
Monomer 839 aa Monomer 1186 aa Dimer 760 aa
NH2 COOH
NH2

COOH
COOH NH2

Dimer 1343 aa Monomer 1400 aa

NH2 NH2
Receptor have extracellular ligand binding
domain, trnas-Membrane and cytoplasmic
Domains.

Receptor could be different Size.

May be monomer or dimer.

Orientation may be different.

COOH COOH
are defective?
Familial hypercholesterolemia
• A genetic defect in LDL
receptors prevents them
from binding to Adaptin-2 .
Familial hypercholesterolemia is a genetic disorder. It
• Thus, they do not enter
is caused by a defect on chromosome 19.
clathrin
Familialcoated pits and can
hypercholesterolemia (FH) is caused by a
mutations
not in into
be brought the LDLR
the cellgene.
via
The defect makes the body unable to remove low
receptor mediated endocytosis.
density lipoprotein (LDL, or bad) cholesterol from the
• The result is high serum
blood. This results in a high level of LDL in the blood.
This makesbecause
cholesterol, you more likely to have narrowing of the
this is
arteries
the from atherosclerosis
mechanism for reducing at an early age.
cholesterol levels and getting
it into the cells.

Low density lipoprotein (LDL) receptor was isolated from a patient with familial
hypercholesterolemia whose receptor fails to cluster in coated pits.

The DNA sequence revealed a substitution of a cysteine codon for a tyrosine codon at
residue 807 in the cytoplasmic domain of the receptor.
 Internalization
signals
LDLR (790-811): Asn-Pro-X-Tyr (Tyr 807)
Surface-exposed tight turns
TFR (19-28) :Tyr-X-Arg-Phe (Tyr20)
generated by Tetrapeptide analogues
Man-6P-R (23-234):Tyr-X-Tyr-X-Lys-Val (Tyr24 & 26) is the general conformational
Insulin R (954-965):Asn-Pro-X-Tyr recognition motif for endocytosis.
EGFR (973-991):Tyr-Arg-Ala-Leu (Tyr974) Curr Opin Cell Biol (1991) 3:634-41
Temperature affects shape of clathrin coated pits
 Clathrin assembly &
Clathrin triskelion invagination

Triskelia assemble as
a array of hexagons on
plasma membrane

Conversion of hexagon to Further conversions

pentagons leads to invagination result in spherical vesicles
 Coated vesicle & Clathrin

Purified coated vesicle treated with

Triskelion 0.5M Tris (pH 7.0) and 2 M Urea
removes three protein of 180,000 Da
(heavy chain), and twopolypeptides
of 36,000 and 33,000 Da (light chain).
These proteins can self assembles into
basket like structure of clathrin when
incubated at pH6.0.

A 100,000 Da protein still associated

With the vesicle after these treatment.
 Domain of the Triskelion

Digestion of cages with

Trypsin cuts the distal
elastase selectively proteolyses
portion of the HC, but
LC. Elastase-digested cages
still retain the ability to
are unable to reassociate into
form cages suggests
cages indicating that LC pull
that central domain of
the distal portion of HC into
HC require for cage
correct position during cage
formation.
assembly.

In a triskelion, the heavy chains interact with each other in the

central hub and protrude outwards as long legs. These legs interact
with other triskelia to form polygonal lattices, the structural
backbone of the clathrin coat.
Formation of Clathrin cage
 Identification of Adaptor

Triskelion

+
Uncoated vesicles with Clathrin Coated vesicles
110,000 Da protein

+
Elastase (remove 110,000 Da)
digested uncoated vesicles
 Adaptors

The term “adaptin” was coined by Barbara Pearse (1975) to

designate a group of 100 kDa proteins that copurified with
clathrin upon isolation of clathrin-coated vesicles.

The 100 kDa-proteins were later found to be subunits of

heterotetrameric adaptor protein (AP) complexes,.

Four basic AP complexes have been described: AP-1, AP-

2, AP-3, and AP-4.

Each of these complexes is composed of two large

adaptins (one each of γ, α, δ or ε and β 1–4, respectively,
90–130 kDa), one medium adaptin (μ1–4, 50 kDa), and one
small adaptin (σ1–4, 20 kDa).
Clathrin coat formation in different locations

AP-2 mediates rapid endocytosis from the

plasma membrane.

AP-1 mediate sorting events at the Golgi

network.

AP-3 mediate sorting events at the trans-

Golgi network (TGN).

AP-4 is most likely part of a nonclathrin coat.

Action of dyanamin in endocytosis

GTP hydrolysis

GTP g S
Receptor mediated endocytosis
References
1. CAMPBELL B I O L O G Y (9th Edition)
Chapter 7
Jane B. ReeceLisa A. Urry
Michael L. Cain
Steven A. Wasserman
Peter V. Minorsky
Robert B. Jackson

2. Molecular Biology of Cells (5th Edition)

Chapter 13
BruceA lberts
AlexandeJro hnson
JulianL ewis
Martin Raff
Keith Roberts
PeteWr alter