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dc.contributor.authorHan, Xinbing
dc.contributor.authorTachado, Souvenir D.
dc.contributor.authorKoziel, Henryk
dc.contributor.authorBoisvert, William A.
dc.date.accessioned2013-03-29T18:31:21Z
dc.date.issued2012
dc.identifier.citationHan, Xinbing, Souvenir D. Tachado, Henry Koziel, and William A. Boisvert. 2012. \(Leu128^{3.43}\) (L128) and \(Val247^{6.40}\) (V247) of CXCR1 are critical amino acid residues for G protein coupling and receptor activation. PLoS ONE 7(8): e42765.en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:10476710
dc.description.abstractCXCR1, a classic GPCR that binds IL-8, plays a key role in neutrophil activation and migration by activating phospholipase C (PLC)β through \(G\alpha_{15}\) and \(G\alpha_i\) which generates diacylglycerol and inositol phosphates (IPs). In this study, two conserved amino acid residues of CXCR1 on the transmembrane domain (TM) 3 and TM6, \(Leu128^{3.43}\) (L128) and \(Val247^{6.40}\) (V247), respectively, were selectively substituted with other amino acids to investigate the role of these conserved residues in CXCR1 activation. Although two selective mutants on Leu128, Leu128Ala (L128A) and Leu128Arg (L128R), demonstrated high binding affinity to IL-8, they were not capable of coupling to G proteins and consequently lost the functional response of the receptors. By contrast, among the four mutants at residue Val247 (TM6.40), replacing Val247 with Ala (V247A) and Asn (V247N) led to constitutive activation of mutant receptors when cotransfected with \(G\alpha_{15}\). The V247N mutant also constitutively activated the \(G\alpha_i\) protein. These results indicate that L128 on TM3.43 is involved in G protein coupling and receptor activation but is unimportant for ligand binding. On the other hand, V247 on TM6.40 plays a critical role in maintaining the receptor in the inactive state, and the substitution of V247 impaired the receptor constraint and stabilized an active conformation. Functionally, there was an increase in chemotaxis in response to IL-8 in cells expressing V247A and V247N. Our findings indicate that \(Leu128^{3.43}\) and \(Val247^{6.40}\) are critical for G protein coupling and activation of signaling effectors, providing a valuable insight into the mechanism of CXCR1 activation.en_US
dc.language.isoen_USen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofdoi:10.1371/journal.pone.0042765en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3427349/pdf/en_US
dash.licenseLAA
dc.subjectBiologyen_US
dc.subjectBiochemistryen_US
dc.subjectProteinsen_US
dc.subjectProtein Structureen_US
dc.subjectTransmembrane Proteinsen_US
dc.subjectComputational Biologyen_US
dc.subjectMacromolecular Structure Analysisen_US
dc.subjectImmunologyen_US
dc.subjectImmune Cellsen_US
dc.subjectMolecular Cell Biologyen_US
dc.subjectSignal Transductionen_US
dc.subjectSignaling in Cellular Processesen_US
dc.subjectG-Protein Signalingen_US
dc.subjectCell Adhesionen_US
dc.title\(Leu128^{3.43}\) (L128) and \(Val247^{6.40}\) (V247) of CXCR1 Are Critical Amino Acid Residues for G Protein Coupling and Receptor Activationen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalPLoS ONEen_US
dash.depositing.authorKoziel, Henryk
dc.date.available2013-03-29T18:31:21Z
dc.identifier.doi10.1371/journal.pone.0042765*
dash.contributor.affiliatedTachado, Souvenir
dash.contributor.affiliatedHan, Xinbing
dash.contributor.affiliatedKoziel, Henryk


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