REPRODUCTION

REVIEW

Focus on Gonadotrophin Signalling

Specificity and promiscuity of gonadotropin receptors
Sabine Costagliola, Eneko Urizar, Fernando Mendive and Gilbert Vassart
Institut de Recherche Interdisciplinaire (IRIBHM) and Department of Medical Genetics, Universite Libre
de Bruxelles, Campus Erasme, 808 route de Lennik, B-1070 Bruxelles, Belgium
Correspondence should be addressed to G Vassart; Email: gvassart@ulb.ac.be

Abstract
The dichotomy between hormone recognition by the ectodomain and activation of the G protein by the rhodopsin-like
serpentine portion is a well established property of glycoprotein hormone receptors. The specificity barrier avoiding
promiscuous activation of the FSH receptor by the high concentration of human chorionic gonadotropin (hCG) prevailing
during human pregnancy was thus believed to lie in the ectodomain. In the past two years, mutations responsible for rare
spontaneous cases of ovarian hyperstimulation syndromes have partially modified this simple view. Five naturally occurring
mutations have been identified which cause an increase in the sensitivity of the FSH receptor to hCG. Surprisingly, these
mutations are all located in the serpentine portion of the receptor. In addition to their effect on sensitivity to hCG, they
increase sensitivity of the FSH receptor to TSH, and are responsible for activating the receptor constitutively. Together, the
available information indicates that the ectodomain and the serpentine domain of the FSH receptor each contribute to the
specificity barrier preventing its spurious activation by hCG. While the former is responsible for establishment of binding
specificity, the latter introduces a novel notion of functional specificity.
Recent data demonstrate that LH and FSH receptors can constitute functional homo- and heterodimers. This suggests the
possibility that in cells co-expressing the two receptors, such as granulosa cells, the heterodimers might be endowed with
functional characteristics different from those of each homodimer.
Reproduction (2005) 130 275281

Introduction The bipartite structure of GpHRs and LGRs is

Amongst the very large family of rhodopsin-like G
protein-coupled receptors (GPCRs), glycoprotein hormone
receptors (GpHRs) constitute a three-member subgroup
made up of the follitropin receptor (FSHR) (Dias et al.
2002), the lutropin receptor (LH/CGR) (Ascoli et al. 2002)
and the thyrotropin receptor (TSHR) (Szkudlinski et al.
2002). These are themselves part of a subfamily of
receptors characterized by an ectodomain containing leu-
cine-rich repeat motifs (LRRs), in addition to the canonical
heptahelical serpentine domain typical of GPCRs. They
are called leucine repeat-containing receptors (LGRs) (Hsu
et al. 2000) and contain the recently identified relaxin
(LGR7) (Hsu et al. 2002) and insulin-like 3 receptors
(LGR8) (Kumagai et al. 2002) together with the receptor
to the insect melanizing hormone bursicon (DLGR2) (Luo
et al. 2005, Mendive et al. 2005) and orphan receptors.
accompanied by a functional dichotomy: their LRR-
containing ectodomain is responsible for the specificity of
binding of their respective agonists, which translates into
activation of the rhodopsin-like serpentine domain, itself
responsible for transducing the signal within the cell,
mainly via activation of the G protein Gs. According to a
model elaborated initially for the TSHR (Vlaeminck et al.
2002), and later extended to the GpHR family (Vassart
et al. 2004, Karges et al. 2005), binding of the hormone to
the receptor would trigger a conformational change in a
motif of the ectodomain, transforming it into an agonist of
the serpentine domain. This model does not require the
postulation of a direct interaction of the agonist with
the extracellular loops or transmembrane helices of the
serpentine domain in order to trigger activation, which
has implications for the understanding of the mechanisms
of inappropriate stimulation of the FSHR by chorionic

q 2005 Society for Reproduction and Fertility DOI: 10.1530/rep.1.00662

ISSN 14701626 (paper) 17417899 (online) Online version via www.reproduction-online.org
276 S Costagliola and others

gonadotropin in spontaneous ovarian hyperstimulation are likely encoded in the serpentine domains (e.g. see
syndrome (OHSS) (see below). Nechamen et al. 2004).

Co-evolution of glycoprotein hormones and their
receptors
Glycoprotein hormones are dimers with a common alpha
subunit and hormone-specific beta subunits encoded by
paralogous genes (i.e. descending from a common ances-
tor). This explains why the beta subunits of thyrotropin
(TSH), luteinizing hormone (LH) and follicle-stimulating
hormone (FSH) share about 40% sequence identity (see
Fig. 1). The corresponding receptors are also encoded by
paralogous genes and, accordingly, they also display
about 40% sequence identity in their hormone-binding
ectodomain (Fig. 1). This fits nicely with the notion that the
hormoneectodomain couples experienced co-evolution,
resulting in tight binding specificity and avoiding pro-
miscuous cross-signaling between the three endocrine sys-
tems (Moyle et al. 1994). In contrast, the serpentine
effector domains of the three receptors share higher
sequence identity (about 70%), which suggests that they
fulfil essentially the same function activation of Gs.
Whereas this conclusion is supported by domain swap-
ping experiments (Braun et al. 1991), it must be kept in
mind that additional signaling functions specific to each
receptor, and distinct from the mere activation of Gs,
Chorionic gonadotropin in primates challenges the
specificity of the system
The specificity barriers between the TSHTSHR, FSHFSHR
and LHLHR hormone receptor couples is such that no
cross-signaling occurs under physiological conditions in
which hormone concentrations are low (i.e. in the picomo-
lar range). In diseases, however, the situation may be differ-
ent. For instance, the extremely high TSH values observed in
congenital hypothyroidism have been implicated in cases of
precocious puberty as the consequence of promiscuous acti-
vation of the FSHR by TSH (Anasti et al. 1995). It is not
widely appreciated that chorionic gonadotropin (CG),
which achieves high concentrations during human preg-
nancy, is a relatively recent invention of evolution. Except
for the equidae, which have also evolved a chorionic gon-
adotropin (Murphy & Martinuk 1991), CG appeared in pri-
mates in which its concentration (and, likely, its role)
increased to reach high nanomolar circulating concen-
trations during the first trimester of human pregnancy. The
beta subunit of human CG (hCG) is 80% identical to beta
LH (hCG and LH stimulate the same LH/CG receptor), and
the disparity between the serum concentrations of TSH, FSH
and LH on the one hand, and CG on the other, constitutes a

NH2

Ectodomain

Serpentine
domain LH/CGr bhCG
Endodomain NH2
COOH
80%

bLH

45% 46%
NH2 NH2
COOH
70% 72% 43% 41%
39%

68% bTSH 40% bFSH

COOH COOH
TSHr FSHr

Receptors Beta subunits of hormones

Figure 1 The beta subunits of the glycoprotein hormones and the glycoprotein hormone receptors are encoded by paralogous genes. Sequence
identities are indicated separately for the ectodomains and the serpentine domains of the three receptors (left), and for the beta subunits of the
four hormones (right). The pattern of shared similarities suggests co-evolution of the hormones and the ectodomain of their receptors, resulting
in the generation of specificity barriers. The high similarity displayed by the serpentine portions of the receptors is compatible with a conserved
mechanism of intramolecular signal transduction. Adapted from Vassart et al. (2004).

Reproduction (2005) 130 275281 www.reproduction-online.org


real challenge to the specificity barriers during the first tri-
mester of human pregnancy. This is readily illustrated by the
inverse relation between hCG and TSH concentrations
observed during this period of normal pregnancy, demon-
strating that hCG reaches concentrations at which it displays
thyrotropic activity (Glinoer 1997). Positively selected
mutations have been identified in the alpha subunits of gly-
coprotein hormones of primates, resulting in lower potency
of the complete hormones when compared with other mam-
mals (Grossmann et al. 1997). This may be interpreted as a
sign of balanced evolution between the negative effects
which would result from cross-signaling (hyperthyroidism,
ovarian hyperstimulation syndrome) and the still hypotheti-
cal positive effects of the high concentrations of hCG. These
may be independent of the LH/CG receptor, since the con-
centrations achieved are well beyond saturation of its classi-
cal concentration action curve (but see below).
The binding specificity of GpHRs is clearly encoded
in their amino terminal ectodomain
From early experiments in which the ectodomains of the
LH and FSH receptors were exchanged (Braun et al.
1991), it is widely accepted that the LRRs of the ectodo-
main are the structures implicated in binding specificity.
In addition, it has convincingly been demonstrated that
purified ectodomains, truncated from their serpentine
domains, do bind their cognate hormones with high affi-
nity (Cornelis et al. 2001, Remy et al. 2001, Schmidt et al.
2001). Very recently, crystallization of a complex between
FSH and the ectodomain of the FSHR has provided the
first direct structural evidence for the residues implicated
in recognition specificity at the atomic level of resolution
(Fan & Hendrickson 2005). Extensive site-directed muta-
genesis experiments, piloted by molecular modeling of
the ectodomains, had previously identified key residues
involved in the specificity of recognition of the TSH and
FSH receptors. The observation that the specificity barriers
could be virtually abolished by exchanging a very limited
number of amino acids between the ectodomain of the
TSHR (8 residues) or the FSHR (2 residues), and the
LH/CGR demonstrated unambiguously that specificity is
encoded in the portion of the ectodomains containing leu-
cine-rich repeats (Smits et al. 2003a) (Fig. 2).
A nice illustration of this conclusion is provided by a
natural mutation of the TSHR rendering it abnormally sen-
sitive to hCG (Rodien et al. 1998). The mutation was
identified in two women who experienced severe
hyperthyroidism on the occasion of each of their pregnan-
cies. It affects a residue, lysine in position 183, predicted
to lie in the sixth LRR, on the hormone-binding surface of
the TSHR. A convincing molecular explanation for the
phenotype was reached by a combination of molecular
modeling and site-directed mutagenesis experiments
(Smits et al. 2002).
www.reproduction-online.org
Specificity and promiscuity of gonadotropin receptors 277
As a logical extension of these notions, mutations
which would make the FSHR abnormally sensitive to hCG
were searched for in spontaneous ovarian hyperstimula-
tion syndromes. It was known that exceptionally high
circulating levels of hCG, as observed for example in
molar pregnancies, are capable of causing ovarian hyper-
stimulation via promiscuous activation of the FSHR
(Ludwig et al. 1998). When FSHR genes were sequenced
from patients displaying recurrent spontaneous OHSS,
mutations were indeed found (Smits et al. 2003b, Vasseur
et al. 2003, Montanelli et al. 2004a, 2004b) but, surpris-
ingly, in all cases studied to date, the mutations affected
residues in the serpentine domain (Fig. 3).
Functional specificity of the FSHR is encoded, in
part, in the serpentine domain
The unexpected observation that mutations of the FSHR
associated with spontaneous OHSS were located in the
serpentine domain of the receptor prompted a detailed
analysis of the phenotype of the mutants, when expressed
by transfection of complementary DNA in cell lines. After
an initial period of disagreement on some aspects of the
phenotype (points 2 and 4 below; Smits et al. 2003b,
Vasseur et al. 2003), we can summarize the characteristics
of these natural mutant receptors in the following way:
(1) they display increased sensitivity to hCG; (2) they also
display increased sensitivity to TSH; (3) they keep normal
sensitivity to FSH; and (4) in contrast to the wild-type
FSHR, which is totally silent, they display detectable con-
stitutive activity (Smits et al. 2003b, Montanelli et al.
2004a, 2004b). The location of the mutations in the top-
ology of the receptor (Fig. 3), together with the obser-
vation that the loss of specificity affected the response to
both hCG and TSH, led to the hypothesis that the under-
lying mechanism might be related to an increased ability
of the mutants to be activated, rather than to an increase
in binding affinity. This hypothesis was directly supported
by experimental evidence: artificial mutations with an
effect on the basal activity of the FSHR, whether they
affect the residues implicated in spontaneous OHSS or
not, caused a similar loss of specificity for hCG and TSH
(Montanelli et al. 2004b). In addition, there was a direct
relation between basal activity of the mutants and the
extent of their sensitivity to the promiscuous hormones
(De Leener A, Montanelli L, Van Durme J, Chae H, Smits
G, Vassert G and Costagliola S, unpublished obser-
vations). From these data, it has been proposed that the
specificity barrier against activation of the FSHR by hCG
evolved by locking the serpentine portion of the receptor
in a completely silent (inactive) state (Vassart et al. 2004).
As such, the wild-type receptor would be unable to
respond to low affinity interaction of hCG (or TSH) with
its ectodomain. In OHSS mutants, the intramolecular bar-
rier to activation would be lower, thus allowing even poor
agonists (such as hCG or TSH) to become effective (Fig.
Reproduction (2005) 130 275281
278 S Costagliola and others

LH/CGr K179G/K104N FSHr

K179G K104N
30
cAMP pmoles/ml

20

10

0
0 103 102 101 100 101 102 103
rhCG IU/ml

Figure 2 Illustration of the specificity barrier encoded in the ectodomain of the FSH receptor. Substitution of two residues of the ectodomain of
the FSHR (K104N, K179G) increases dramatically its response to hCG. The left panel shows responsiveness to hCG in terms of cAMP production
of the wild-type FSHR (black circles), the K104N (green diamonds), the K179G (blue circles) and the double K179G/K104N mutant (red circles)
after transfection in COS-7 cells. The responsiveness of the wild-type LH/CG receptor is shown for comparison (black squares). The right panel
illustrates the position of K104 and K179 on the tridimensional structure of the ectodomain of the FSHR (Fan & Hendrickson 2005).

Figure 3 Schematic representation of the FSH receptor, with indication of the mutations identified in patients with spontaneous ovarian hypersti-
mulation syndrome: D6.30N (Smits et al. 2003b); D6.30G (De Leener et al., unpublished observations; see page 3); T3.32I (Vasseur et al. 2003);
T3.32A (Montanelli et al. 2004a); I5.54T (De Leener et al., unpublished observations). The standardized numbering system used to identify
amino acids is from Ballesteros & Weinstein (2002).

4). In favor of such an interpretation, one of the residues
affected by OHSS mutation (D6.30) (see Fig. 3) has been
identified as a key player in the maintenance of rhodop-
sin-like GPCRs in the inactive state (Ballesteros et al.
2001). Mutations of the homolog residues in the TSHR or
LH/CGR cause constitutive activation responsible for
hyperfunctioning thyroid adenomas and autosomal domi-
nant hyperthyroidism (Parma et al. 1993, Duprez et al.
1994), or male-limited pseudoprecocious puberty (Shen-
ker et al. 1993, Themmen & Huhtaniemi 2000)
respectively.
If the above hypothesis is correct, one might expect
non-primate FSHRs to display some sensitivity to hCG,
when tested ex vivo, together with some basal activity.
This is indeed the case, as suggested by the phenotype of
chimeric receptors containing the ectodomain of the
human FSHR pasted upstream of a serpentine domain
from a series of non-primates species (De Leener et al.,
unpublished observations). Most chimeras displayed
increased sensitivity to hCG when compared with the
wild-type human FSHR. Interestingly, the relation between
basal activity and relaxed specificity seems to be a charac-
teristic of the FSHR. Mutants of the TSHR endowed with
increased constitutive activity do not show a significant
increase in sensitivity to hCG (Montanelli et al. 2004b). It
must be noted, however, that contrary to the FSHR, the
wild-type human TSHR exhibits easily measurable consti-
tutive activity (Parma et al. 1993). It seems, therefore, that

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 Specificity and promiscuity of gonadotropin receptors 279

Figure 4 The serpentine domain of the FSH receptor participates in its functional specificity. The receptors are represented with their ectodomain
containing a concave, hormone binding structure facing upwards, and a transmembrane serpentine portion. Our current interpretation of the
phenotypes displayed by the FSHR mutants of patients with spontaneous ovarian hyperstimulation syndrome (OHSS) is depicted. High affinity
interactions of the ectodomain of the wild-type (WT) FSHR (FSHr) with FSH (upper path, A) would result in full activation of the serpentine
portion (B). High concentrations of human chorionic gonadotropin (hCG), while capable of establishing low affinity interactions with the
ectodomain of the FSHR (lower path) would be ineffective in activating a strongly locked wild-type (wt) serpentine domain (C). In OHSS mutants
(mutation D6.30N illustrated by a red dot), partial activation of the serpentine domains (indicated as a partially relaxed structure, in blue) would
lower the activation threshold of the serpentine, thus allowing stimulation to take place (D).

evolution has followed distinct paths in the TSHR and
FSHR to cope with the emergence of chorionic
gonadotropin. In the former, maintenance of specificity
relied on evolution of the binding barrier within the ecto-
domain. In the latter, an intramolecular barrier to acti-
vation was increased by strengthening silencing locks of
the serpentine domain.
Additional complexity from dimerization of
receptors
Over the past few years, the notion that GPCRs are present
in the plasma membrane as dimers or oligomers has pro-
gressively become the prevalent view (Fotiadis et al. 2003,
Terrillon & Bouvier 2004). In addition, the possibility that
receptors belonging to the same or close subfamilies can
associate into heterodimers has been convincingly docu-
mented. The GpHRs are no exception (Osuga et al. 1997,
Horvat et al. 2001, Ji et al. 2002, Latif et al. 2002, Tao et al.
2004, Urizar et al. 2005). There were few arguments,
however, for a possible functional significance of homo- or
hetero-dimerization of GpHRs until recently, when it was
shown that dimerization was associated with allostery, more
precisely with negative cooperativity (Urizar et al. 2005).
Briefly stated, these novel data contend that GpHRs are
present in the plasma membrane as homodimers and that
binding of the hormone to one subunit of the dimer
decreases strongly the affinity of the other for the ligand.
When two different GpHRs are present in the same cell,
transfection experiments showed that they are capable of
forming heterodimers, and these also display negative coop-
erativity. This means that in cells co-expressing FSHR and
LH/CGR, as granulosa cells do before luteinization (Hillier
2001), the possibility exists of an interplay between the two
hormones at the level of putative receptor heterodimers.
One consequence of these observations is that under most
physiological conditions (i.e. at low agonist concentration) a
single agonist would be bound per dimer. However, given
the high concentration of hCG achieved during pregnancy,
we must consider the possibility that dimers (be they homo-
or hetero-dimers) could show different abilities to activate
downstream regulatory cascades when they are engaged by
a single or by two molecules of agonist.
Perspectives
It is hoped that the recent structural and functional data
collected on gonadotropins and their receptors will open
new perspectives for understanding the pathophysiology
of a series of diseases, in addition to providing new

www.reproduction-online.org Reproduction (2005) 130 275281

280 S Costagliola and others
insights into the basic mechanisms involved in their
action. In particular, we believe that the following points
warrant further investigation: (1) the putative role of FSHR
variants in the severity of iatrogenic OHSS (Daelemans
et al. 2004); (2) the role of putative FSHR/LHR heterodi-
mers in granulosa cells at the time of ovulation; and (3)
the putative role of LHR dimers as the target for the high,
saturating concentrations of hCG during pregnancy.
Acknowledgements
We thank Veronique Janssens and all the staff of the Costa-
gliola laboratory for their contribution to the experimental
work summarized in this review. Our gratitude goes to the
clinicians who referred the OHSS cases (Drs O Olatunbosun,
R Pierson, C Di Carlo, C Nappi and H Chae) and to the
patients themselves. S C is Chercheur Qualifie at the FNRS.
This study was supported by the Belgian program on Interuni-
versity Poles of Attraction initiated by the Belgian State,
Prime Ministers Office, Science Policy Programming and the
LifeSci Health program of the European Community (grant
LSHB-CT-2003-503337). The work was also supported by
grants from FRSM, FNRS and ARBD. The authors declare that
there is no conflict of interest that would prejudice the impar-
tiality of this scientific work
References
Anasti JN, Flack MR, Froehlich J, Nelson LM & Nisula BC 1995 A
potential novel mechanism for precocious puberty in juvenile
hypothyroidism 8. Journal of Clinical Endocrinology and
Metabolism 80 276279.
Ascoli M, Fanelli F & Segaloff DL 2002 The lutropin/choriogonado-
tropin receptor, a 2002 perspective. Endocrine Reviews 23
141174.
Ballesteros JA & Weinstein SP 2002 Integrated methods for the
construction of three-dimensional models and computational
probing of structurefunction relations in G-protein-coupled
receptors. In Methods in Neurosciences, pp 366 389. Ed. SC
Sealfon. San Diego; Academic Press.
Ballesteros JA, Jensen AD, Liapakis G, Rasmussen SG, Shi L, Gether
U & Javitch JA 2001 Activation of the beta 2-adrenergic receptor
involves disruption of an ionic lock between the cytoplasmic
ends of transmembrane segments 3 and 6. Journal of Biological
Chemistry 276 2917129177.
Braun T, Schofield PR & Sprengel R 1991 Amino-terminal leucine-
rich repeats in gonadotropin receptors determine hormone
selectivity. EMBO Journal 10 18851890.
Cornelis S, Uttenweiler-Joseph S, Panneels V, Vassart G & Costa-
gliola S 2001 Purification and characterization of a soluble
bioactive amino-terminal extracellular domain of the human
thyrotropin receptor. Biochemistry 40 98609869.
Daelemans C, Smits G, de Maertelaer V, Costagliola S, Englert Y,
Vassart G & Delbaere A 2004 Prediction of severity of symptoms
in iatrogenic ovarian hyperstimulation syndrome by follicle-stimu-
lating hormone receptor Ser680Asn polymorphism. Journal of
Clinical Endocrinology and Metabolism 89 63106315.
Dias JA, Cohen BD, Lindau-Shepard B, Nechamen CA, Peterson AJ
& Schmidt A 2002 Molecular, structural, and cellular biology of
follitropin and follitropin receptor. Vitamins and Hormones 64
249322.
Duprez L, Parma J, Van Sande J, Allegeier A, Lecle`re J, Schvartz C,
Delisle MJ, Decoulx M, Orgiazzi J, Dumont JE & Vassart G 1994
Germline mutations in the thyrotropin receptor gene cause non
Reproduction (2005) 130 275281
autoimmune autosomal dominant hyperthyroidism. Nature
Genetics 7 396 401.
Fan QR & Hendrickson WA 2005 Structure of human follicle-stimu-
lating hormone in complex with its receptor. Nature 433
269277.
Fotiadis D, Liang Y, Filipek S, Saperstein DA, Engel A & Palczewski K
2003 Atomic-force microscopy: rhodopsin dimers in native disc
membranes. Nature 421 127128.
Glinoer D 1997 The regulation of thyroid function in pregnancy:
pathways of endocrine adaptation from physiology to pathology.
Endocrine Reviews 18 404 433.
Grossmann M, Weintraub BD & Szkudlinski MW 1997 Novel
insights into the molecular mechanisms of human thyrotropin
action: structural, physiological, and therapeutic implications for
the glycoprotein hormone family. Endocrine Reviews 18 476501.
Hillier SG 2001 Gonadotropic control of ovarian follicular growth
and development. Molecular and Cellular Endocrinology 179
3946.
Horvat RD, Barisas BG & Roess DA 2001 Luteinizing hormone
receptors are self-associated in slowly diffusing complexes during
receptor desensitization. Molecular Endocrinology 15 534542.
Hsu SY, Kudo M, Chen T, Nakabayashi K, Bhalla A, van der Spek PJ,
van Duin M & Hsueh AJ 2000 The three subfamilies of leucine-
rich repeat-containing G protein-coupled receptors (LGR):
identification of LGR6 and LGR7 and the signaling mechanism for
LGR7. Molecular Endocrinology 14 12571271.
Hsu SY, Nakabayashi K, Nishi S, Kumagai J, Kudo M, Sherwood OD
& Hsueh AJ 2002 Activation of orphan receptors by the hormone
relaxin. Science 295 671 674.
Ji I, Lee C, Song Y, Conn PM & Ji TH 2002 Cis- and trans-activation
of hormone receptors: the LH receptor. Molecular Endocrinology
16 12991308.
Karges B, Gidenne S, Aumas C, Haddad F, Kelly PA, Milgrom E &
De Roux N 2005 Zero-length cross-linking reveals that tight inter-
actions between the extracellular and transmembrane domains of
the LH receptor persist during receptor activation. Molecular
Endocrinology 19 20862098.
Kumagai J, Hsu SY, Matsumi H, Roh JS, Fu P, Wade JD, Bathgate RA
& Hsueh AJ 2002 INSL3/Leydig insulin-like peptide activates the
LGR8 receptor important in testis descent. Journal of Biological
Chemistry 277 31283 31286.
Latif R, Graves P & Davies TF 2002 Ligand-dependent inhibition of
oligomerization at the human thyrotropin receptor. Journal of Bio-
logical Chemistry 277 4505945067.
Ludwig M, Gembruch U, Bauer O & Diedrich K 1998 Ovarian
hyperstimulation syndrome (OHSS) in a spontaneous pregnancy
with fetal and placental triploidy: information about the general
pathophysiology of OHSS. Human Reproduction 13 20822087.
Luo CW, Dewey EM, Sudo S, Ewer J, Hsu SY, Honegger HW &
Hsueh AJ 2005 Bursicon, the insect cuticle-hardening hormone, is
a heterodimeric cystine knot protein that activates G protein-
coupled receptor LGR2. PNAS 102 28202825.
Mendive FM, Van Loy T, Claeysen S, Poels J, Williamson M, Hauser F,
Grimmelikhuijzen CJ, Vassart G & Vanden Broeck J 2005
Drosophila molting neurohormone bursicon is a heterodimer and
the natural agonist of the orphan receptor DLGR2. FEBS Letters
579 2171 2176.
Montanelli L, Delbaere A, Di Carlo C, Nappi C, Smits G, Vassart G
& Costagliola S 2004a A mutation in the follicle-stimulating
hormone receptor as a cause of familial spontaneous ovarian
hyperstimulation syndrome. Journal of Clinical Endocrinology and
Metabolism 89 1255 1258.
Montanelli L, Van Durme JJ, Smits G, Bonomi M, Rodien P, Devor
EJ, Moffat-Wilson K, Pardo L, Vassart G & Costagliola S 2004b
Modulation of ligand selectivity associated with activation of
the transmembrane region of the human follitropin receptor.
Molecular Endocrinology 18 2061 2073.
Moyle WR, Campbell RK, Myers RV, Bernard MP, Han Y & Wang X
1994 Co-evolution of ligand-receptor pairs. Nature 368 251 255.
www.reproduction-online.org

Murphy BD & Martinuk SD 1991 Equine chorionic gonadotropin.
Endocrine Reviews 12 2744.
Nechamen CA, Thomas RM, Cohen BD, Acevedo G, Poulikakos PI,
Testa JR & Dias JA 2004 Human follicle-stimulating hormone
(FSH) receptor interacts with the adaptor protein APPL1 in HEK
293 cells: potential involvement of the PI3K pathway in FSH
signaling. Biology of Reproduction 71 629636.
Osuga Y, Hayashi M, Kudo M, Conti M, Kobilka B & Hsueh AJ 1997
Co-expression of defective luteinizing hormone receptor fragments
partially reconstitutes ligand-induced signal generation. Journal of
Biological Chemistry 272 2500625012.
Parma J, Duprez L, Van Sande J, Cochaux P, Gervy C, Mockel J,
Dumont JE & Vassart G 1993 Somatic mutations in the thyrotropin
receptor gene cause hyperfunctioning thyroid adenomas. Nature
365 649651.
Remy JJ, Nespoulous C, Grosclaude J, Grebert D, Couture L, Pajot E
& Salesse R 2001 Purification and structural analysis of a soluble
human chorionogonadotropin hormone-receptor complex. Journal
of Biological Chemistry 276 16811687.
Rodien P, Bremont C, Sanson ML, Parma J, Van Sande J, Costagliola
S, Luton JP, Vassart G & Duprez L 1998 Familial gestational
hyperthyroidism caused by a mutant thyrotropin receptor
hypersensitive to human chorionic gonadotropin. New England
Journal of Medicine 339 1823 1826.
Schmidt A, MacColl R, Lindau-Shepard B, Buckler DR & Dias JA
2001 Hormone-induced conformational change of the purified
soluble hormone binding domain of follitropin receptor
complexed with single chain follitropin. Journal of Biological
Chemistry 276 2337323381.
Shenker A, Laue L, Kosugi S, Merendino JJ, Minegishi T & Cutler GB
1993 A constitutively activating mutation of the luteinizing
hormone receptor in familial male precocious puberty. Nature 365
652654.
Smits G, Govaerts C, Nubourgh I, Pardo L, Vassart G & Costagliola S
2002 Lysine 183 and glutamic acid 157 of the thyrotropin receptor:
two interacting residues with a key role in determining specificity
towards TSH and hCG. Molecular Endocrinology 16 722735.
Smits G, Campillo M, Govaerts C, Janssens V, Richter C, Vassart G,
Pardo L & Costagliola S 2003a Glycoprotein hormone receptors:
determinants in leucine-rich repeats responsible for ligand
specificity. EMBO Journal 22 2692 2703.
www.reproduction-online.org
Specificity and promiscuity of gonadotropin receptors 281
Smits G, Olatunbosun O, Delbaere A, Pierson R, Vassart G & Costa-
gliola S 2003b Ovarian hyperstimulation syndrome due to a
mutation in the follicle-stimulating hormone receptor. New Eng-
land Journal of Medicine 349 760766.
Szkudlinski MW, Fremont V, Ronin C & Weintraub BD 2002
Thyroid-stimulating hormone and thyroid-stimulating hormone
receptor structurefunction relationships. Physiology Reviews 82
473 502.
Tao YX, Johnson NB & Segaloff DL 2004 Constitutive and agonist-
dependent self-association of the cell surface human lutropin
receptor. Journal of Biological Chemistry 279 59045914.
Terrillon S & Bouvier M 2004 Roles of G-protein-coupled receptor
dimerization. EMBO Reports 5 3034.
Themmen APN & Huhtaniemi IT 2000 Mutations of gonadotropins
and gonadotropin receptors: elucidating the physiology and
pathophysiology of pituitarygonadal function. Endocrine Reviews
21 551 583.
Urizar E, Montanelli L, Loy T, Bonomi M, Swillens S, Gales C,
Bouvier M, Smits G, Vassart G & Costagliola S 2005 Glycoprotein
hormone receptors: link between receptor homodimerization and
negative cooperativity. EMBO Journal 24 19541964.
Vassart G, Pardo L & Costagliola S 2004 A molecular dissection of
the glycoprotein hormone receptors. Trends in Biochemical
Science 29 119126.
Vasseur C, Rodien P, Beau I, Desroches A, Gerard C, de Poncheville
L, Chaplot S, Savagner F, Croue A, Mathieu E, Lahlou N,
Descamps P & Misrahi M 2003 A chorionic gonadotropin-
sensitive mutation in the follicle-stimulating hormone receptor as a
cause of familial gestational spontaneous ovarian hyperstimulation
syndrome. New England Journal of Medicine 349 753 759.
Vlaeminck V, Ho SC, Rodien P, Vassart G & Costagliola S 2002
Activation of the cAMP pathway by the TSH receptor involves
switching of the ectodomain from a tethered inverse agonist to an
agonist. Molecular Endocrinology 16 736 746.
Received 27 May 2005
First decision 1 June 2005
Revised manuscript received 14 June 2005
Accepted 24 June 2005
Reproduction (2005) 130 275281