Insight into Mutation-Induced Activation of the Luteinizing Hormone Receptor: Molecular Simulations Predict the Functional Behavior of Engineered Mutants at M398

Fanelli, Francesca; Verhoef-Post, Miriam; Timmerman, M. A.; Zeilemaker, Annelieke; Martens, John W.M.; Themmen, Axel P. N.

doi:10.1210/me.2003-0050

Cited by 33 publications

(77 citation statements)

References 32 publications

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“…The SAS computed over R464, T467, I468, and K463 is represented by gray dots. This index is 28 Ǻ 2 for WT LHR and 120 Ǻ 2 for the D564G mutant (Fanelli et al, 2004). Comparison of WT LHR and gain-of-function LHR mutants in response to exogenous hCG.…”

Section: Discussionmentioning

confidence: 99%

“…7B) and non-constitutively active LHR (Fig. 7A), respectively (Angelova et al, 2002;Fanelli, et al, 2004;Zhang et al, 2005). The higher SAS values are hypothesized to correspond to an open form of LHR that facilitates productive Gsα binding.…”

Section: Activation Of the Transmembrane Region Of Lhrmentioning

confidence: 97%

“…In the past several years, a great deal has been learned from modeling the endodomain of LHR using ab initio and, more recently, comparative modeling approaches (Fanelli, 2000;Fanelli et al, 2001Fanelli et al, , 2004Fanelli and Puett, 2002;Fanelli and De Benedetti, 2005;Angelova et al, 2002;Ascoli et al, 2002;Zhang et al, 2005).…”

Section: Activation Of the Transmembrane Region Of Lhrmentioning

confidence: 99%

“…With a significant number of characterized engineered and naturally occurring gain-of-function mutation now available (Huhtaniemi and Themmen, 2005), it is interesting to note the variability that exists in function (Fanelli et al, 2001(Fanelli et al, , 2004Angelova et al, 2002;Ascoli et al, 2002;Zhang et al, 2005). Some, for example, are marginally active, others highly so; some are responsive to exogenous hCG, while others are not (Fig.…”

Section: Activation Of the Transmembrane Region Of Lhrmentioning

confidence: 99%

“…While there is considerable physiological and pharmacological interest in the GPCRs, the only structure available is that for rhodopsin (Palczewski et al, 2000). This structure has thus served as a template for modeling LHR (Fanelli et al, 2001(Fanelli et al, , 2004Fanelli and Puett, 2002;Angelova et al, 2002;Ascoli et al, 2002) and other GPCRs.…”

Section: Introductionmentioning

confidence: 99%

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A functional transmembrane complex: The luteinizing hormone receptor with bound ligand and G protein

Puett

DeMars

et al. 2007

Molecular and Cellular Endocrinology

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The luteinizing hormone receptor (LHR) is one of eight members in a cluster of the rhodopsin family of the large G protein-coupled receptor (GPCR) superfamily that contains some 800-900 genes in the human genome. LHR, along with its paralogons, follicle stimulating hormone receptor (FSHR) and thyroid stimulating hormone receptor, form one of the three classes in this cluster; the two other classes contain the relaxin-binding GPCRs and orphan GPCRs. These GPCRs are characterized by a relatively large ectodomain (ECD) containing leucine-rich-repeats (LRRs); in the class of glycoprotein hormone receptors, the LRR region is capped by N-terminal and C-terminal cysteinerich regions. Binding of human chorionic gonadotropin (hCG) or luteinizing hormone to the LHR-ECD triggers a conformational change of the transmembrane region of the receptor facilitating binding and activation of Gs, followed by effector enzyme activation and subsequent intracellular signaling. Viewing LHR as a transmembrane anchoring protein that sequentially binds hCG and Gs to give the hCG-LHR-Gs complex, numerous interactions and conformational changes must be considered. There is, unfortunately, a paucity of structural data on LHR, but crystal structures exist for hCG, the homologous FSH-FSHR-ECD (N-terminal fragment) complex, rhodopsin (in the inactive state), an active form of Gαs (transducin), and the βγ heterodimer. Using a combined experimental (site-directed mutagenesis followed by characterization in transfected cells) and computational (homology modeling and molecular dynamics simulations) approach, good working models are being developed for the protein-protein interaction faces and, in some cases, the ensuing conformational changes induced by complex formation. hCG binding to the LHR-ECD appears to involve several LRRs; LHR activation can be described in terms of disrupting a network of H-bonds in the cytosolic halves of helices 1-3, 6, and 7; and binding of LHR to Gs involves, in large part, intracellular loop 2 binding, presumably to Gsα at its C-terminus. Major gaps exist in our understanding at the molecular level of the six-polypeptide chain complex, hCG-LHR-Gs, but considerable progress has been made in the past few years.

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Section: Discussionmentioning

confidence: 99%

Section: Activation Of the Transmembrane Region Of Lhrmentioning

confidence: 97%

Section: Activation Of the Transmembrane Region Of Lhrmentioning

confidence: 99%

Section: Activation Of the Transmembrane Region Of Lhrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A functional transmembrane complex: The luteinizing hormone receptor with bound ligand and G protein

Puett

DeMars

et al. 2007

Molecular and Cellular Endocrinology

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Regulatory Processes Governing the Cell Surface Expression of LH and FSH Receptors

Segaloff

2012

Subcellular Biochemistry

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The LH receptor (LHR) and FSH receptor (FSHR), collectively termed the gonadotropin receptors, are members of the Family A of GPCRs. The gonadotropin receptors each contain N-linked carbohydrates that are not directly involved in hormone binding, but contribute to the proper folding, and therefore, cell surface expression of the receptor. Loss-of-function mutations of an LHR or FSHR results in decreased target cell responsiveness. Most inactivating mutations cause receptor misfolding, resulting in the retention of the mutant in its immature form in the endoplasmic reticulum. A membrane-permeable allosteric agonist of the LHR has been shown to serve as a pharmacological chaperone for misfolded and intracellularly retained LHRs by promoting their cell surface expression. Wild-type LHR and FSHR each form homodimers and heterodimers while in the ER. Therefore, when wild-type receptor is co-expressed with a misfolded mutant, the misfolded receptor dimerizes with immature wild-type receptor in the ER, causing a dominant-negative effect on cell surface expression of the mature wild-type receptor. Notably, the propensity for homodimerization is not affected by the activation status of the receptor. However, within a receptor dimer, the activity of one protomer may allosterically regulate the other protomer. Therefore, the dimerization of the gonadotropin receptors appears to be an obligate process that is part of the normal itinerary for trafficking to the cell surface and, once there, the dimerized receptors allow for additional modulations of cell signaling.

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Conserved amino acids participate in the structure networks deputed to intramolecular communication in the lutropin receptor

Angelova

Felline

Lee

et al. 2010

Cell. Mol. Life Sci.

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The luteinizing hormone receptor (LHR) is a G protein-coupled receptor (GPCR) particularly susceptible to spontaneous pathogenic gain-of-function mutations. Protein structure network (PSN) analysis on wild-type LHR and two constitutively active mutants, combined with in vitro mutational analysis, served to identify key amino acids that are part of the regulatory network responsible for propagating communication between the extracellular and intracellular poles of the receptor. Highly conserved amino acids in the rhodopsin family GPCRs participate in the protein structural stability as network hubs in both the inactive and active states. Moreover, they behave as the most recurrent nodes in the communication paths between the extracellular and intracellular sides in both functional states with emphasis on the active one. In this respect, non-conservative loss-of-function mutations of these amino acids is expected to impair the most relevant way of communication between activating mutation sites or hormone-binding domain and G protein recognition regions.

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Insight into Mutation-Induced Activation of the Luteinizing Hormone Receptor: Molecular Simulations Predict the Functional Behavior of Engineered Mutants at M398

Cited by 33 publications

References 32 publications

A functional transmembrane complex: The luteinizing hormone receptor with bound ligand and G protein

A functional transmembrane complex: The luteinizing hormone receptor with bound ligand and G protein

Regulatory Processes Governing the Cell Surface Expression of LH and FSH Receptors

Conserved amino acids participate in the structure networks deputed to intramolecular communication in the lutropin receptor

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