Computational Modeling Approaches to Structure−Function Analysis of G Protein-Coupled Receptors

Fanelli, Francesca; Benedetti, Pier G. De

doi:10.1021/cr000095n

Cited by 151 publications

(158 citation statements)

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“…The extensive in silico mutagenesis experiments on naturally occurring as well as artificial single and multiple activating mutations of the hLHR made it possible to infer the structural triggers of mutation-induced receptor activation (15). Two of these mutations (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…Although it was originally hypothesized that the D578 (6.44) H mutant uniquely caused activation of the phospholipase C signaling pathway (8), more recent studies suggest that the G proteins and second messenger pathways activated by * These studies were supported by National Institutes of Health Grant D578 (6.44) H are the same as those activated by other hLHR CAMs (10,11). Computational modeling on the isolated wild type (WT) and constitutively active mutants of the hLHR suggests that ligand-independent activation of the hLHR involves changes in the interaction pattern of Arg-464 (3.50) of the highly conserved (E/D)R(Y/W) motif and the opening of a solventaccessible crevice in the neighbors of this conserved arginine (12)(13)(14)(15).…”

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confidence: 99%

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Intrinsic Differences in the Response of the Human Lutropin Receptor Versus the Human Follitropin Receptor to Activating Mutations

Zhang

Tao

Ryan

et al. 2007

Journal of Biological Chemistry

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In contrast to the human lutropin receptor (hLHR), very few naturally occurring activating mutations of the structurally related human follitropin receptor (hFSHR) have been identified. The present study was undertaken to determine if one aspect underlying this discrepancy might be a general resistance of the hFSHR to mutation-induced constitutive activity. Five different mutations were introduced into both the hLHR and hFSHR (four based on activating mutations of the hLHR gene, one based on an activating mutation of the hFSHR gene). Our results demonstrate that hFSHR constitutively activating mutants (CAMs) were not as active as hLHR CAMs containing the comparable mutation. Furthermore, although all hFSHR CAMs exhibited strong promiscuous activation by high concentrations of the other glycoprotein hormone receptors, hLHR CAMs showed little or no promiscuous activation. Our in vitro findings are consistent with in vivo observations of known pathophysiological conditions associated with hLHR CAMs, but not hFSHR CAMs, and with promiscuous activation of hFSHR CAMs, but not hLHR CAMs. Computational experiments suggest that the mechanisms through which homologous mutations increase the basal activity of the hLHR and the hFSHR are similar. This is particularly true for the strongest CAMs like L460 (3.43) R. Disparate properties of the hLHR versus hFSHR CAMs may, therefore, be due to differences in shape and electrostatics features of the solventexposed cytosolic receptor domains involved in the receptor-G protein interface rather than to differences in the nature of local perturbation at the mutation site or in the way local perturbation is transferred to the putative G protein binding domains. The LH receptor (LHR)4 and FSH receptor (FSHR), collectively termed the gonadotropin receptors, are G protein-coupled receptors whose primary role is mediation of the signal transduction by pituitary LH or placental hCG (LHR) or pituitary FSH (FSHR) in the gonads. The LHR and FSHR are each composed of a serpentine region containing the seven transmembrane helices typical of G protein-coupled receptors as well as a large extracellular domain that confers the high affinity binding of hormone (1, 2). The gonadotropin receptors are members of the leucine-rich glycoprotein receptor subfamily of rhodopsin-like G protein-coupled receptors (3). The gonadotropin receptors of human origin are highly homologous, with the greatest degree of amino acid conservation within the transmembrane helices. Recent crystallographic studies on human FSH (hFSH) bound to the extracellular domain of the hFSHR have tremendously advanced our understanding of the mechanism of hormone binding to the gonadotropin receptors (4). The entire receptor complexed with ligand has yet to be crystallized, however, and it is still unclear how the binding of hormone to the extracellular domain of the gonadotropin receptor causes stabilization of the serpentine region in an active conformation that causes stimulation of G proteins, primarily G s .In the past several year...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Intrinsic Differences in the Response of the Human Lutropin Receptor Versus the Human Follitropin Receptor to Activating Mutations

Zhang

Tao

Ryan

et al. 2007

Journal of Biological Chemistry

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“…An exception may be when significant amounts of experimental information are available to provide constraints for refinement, such as for GPCRs (54).…”

Section: D-structure Predictionmentioning

confidence: 99%

Membrane protein prediction methods

Punta

Forrest

Bigelow

et al. 2007

Methods

109

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We survey computational approaches that tackle membrane protein structure and function prediction. While describing the main ideas that have led to the development of the most relevant and novel methods, we also discuss pitfalls, provide practical hints and highlight the challenges that remain. The methods covered include: sequence alignment, motif search, functional residue identification, transmembrane segment and protein topology predictions, homology and ab initio modeling. Overall, predictions of functional and structural features of membrane proteins are improving, although progress is hampered by the limited amount of high-resolution experimental information available. While predictions of transmembrane segments and protein topology rank among the most accurate methods in computational biology, more attention and effort will be required in the future to ameliorate database search, homology and ab initio modeling.

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“…Efficient coupling of GPCRs to their cognate heterotrimeric G proteins through dimeric or monomeric forms of the receptors has been suggested in modeling studies (3,4), but so far, only a pentameric complex composed of two GPCRs and G protein has been isolated (5). In our model, one rhodopsin (Rho) molecule needs to be activated whereas the second serves as a platform to support G protein binding and does not require activation (3).…”

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confidence: 98%

Functional and Structural Characterization of Rhodopsin Oligomers

Jastrzębska

Fotiadis

Jang

et al. 2006

Journal of Biological Chemistry

127

102

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A major question in G protein-coupled receptor signaling concerns the quaternary structure required for signal transduction. Do these transmembrane receptors function as monomers, dimers, or larger oligomers? We have investigated the oligomeric state of the model G protein-coupled receptor rhodopsin (Rho), which absorbs light and initiates a phototransduction-signaling cascade that forms the basis of vision. In this study, different forms of Rho were isolated using gel filtration techniques in mild detergents, including n-dodecyl-␤-D-maltoside, n-tetradecyl-␤-D-maltoside, and n-hexadecyl-␤-D-maltoside. The quaternary structure of isolated Rho was determined by transmission electron microscopy, demonstrating that in micelles containing n-dodecyl-␤-D-maltoside, Rho exists as a mixture of monomers and dimers whereas in n-tetradecyl-␤-D-maltoside and n-hexadecyl-␤-D-maltoside Rho forms higher ordered structures. Especially in n-hexadecyl-␤-D-maltoside, most of the particles are present in tightly packed rows of dimers. The oligomerization of Rho seems to be important for interaction with its cognate G protein, transducin. Although the activated Rho (Meta II) monomer or dimers are capable of activating the G protein, transducin, the activation process is much faster when Rho exists as organized dimers. Our studies provide direct comparisons between signaling properties of Meta II in different quaternary complexes.

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Computational Modeling Approaches to Structure−Function Analysis of G Protein-Coupled Receptors

Cited by 151 publications

References 628 publications

Intrinsic Differences in the Response of the Human Lutropin Receptor Versus the Human Follitropin Receptor to Activating Mutations

Intrinsic Differences in the Response of the Human Lutropin Receptor Versus the Human Follitropin Receptor to Activating Mutations

Membrane protein prediction methods

Functional and Structural Characterization of Rhodopsin Oligomers

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