---------------------------------------------------------------------------- GPCR TM mutations resulting in constitutive activity (CA) ---------------------------------------------------------------------------- I:9 LHR human L368P -> FMPP incr basal cAMP Latronico et al. J Clin Endo Metab 85:4799 (2000) I:14 LHR human A373V Simoni et al. Steroids 63:288 (1998) I:17 TSHR human G431S incr basal cAMP and IP; C,A incr basal cAMP Biebermann et al. J Clin Endo Metab 86:4429 (2001) I:18 alpha1bAR N63A not D incr basal IP Scheer et al. EMBO J 15:3566 (1996) =-=-=-=-=-=-=-=-=-= II:1 melanocortin1R (MSHR) mouse tobacco darkening mutant S69L Robbins et al. Cell 72:827 (1993) II:3 adenosine A2BR human T42A constit act Li et al. J Pharmacol Exp Ther. 2006 Oct 31 [Epub ahead of print] II:4 KSHV-GPCR D83A,N further increase basal IP's (wt is constit act) Ho et al. JBC 276(2):1376 (2001) II:7 LHR human M398T -> familial male precocious puberty (FMPP) incr basal cAMP Kraaij et al. J Clin Endo Metab 80:3160 (1995) Yano et al. BBRC 220:1036 (1996) Ignacak et al. Endocrine J 47:595 (2000) II:7 TSHR human M453T Parma et al. J Clin Endo Metab 82(8):2695 (1997) II:10 TSHR human L456A not I,W incr expression-normalized basal cAMP Urizar et al. JBC 280(17):17135 (2005) II:14 B1AR human D104A not K Ahmed et al. Life Sci 78:2019 (2006) Hossain et al. Biol Pharm Bull 31:1517 (2008) II:17 TSHR human M->V Smit et al. Annu Rev Pharmacol Toxicol 47:22.1 (2007) II:20 CCR5 human T82K,R>P,H,Y not G,A,V,L,M,W,D,E constit act in yeast assay Alvarez Arias et al. JBC online (2003) II:20 CCR2 human T94K not P constit act in yeast assay Alvarez Arias et al. JBC online (2003) II:21 rhodopsin bovine G90D [postulate subst for Schiff base counterion] Rao et al. Nature 367:639 (1994) II:21 CCKBR human C107A incr basal IP's Blaker et al. Mol Pharm 58:399 (2000) II:24 melanocortin1R (MSHR) mouse E92R,K (not A,D,Q), yet decr ag affinity Lu et al. Mol Endocrinol 12:592 (1998) [postulate agonist-mimetic] Robbins et al. Cell 72:827 (1993) sombre mutation E92K II:25 rhodopsin T94I Inglehearn Eye 12:571 (1998). (II:29) ()=not in Baldwin 3D model melanocortin1R (MSHR) bovine L99P dominant black color Lu et al. Mol Endocrinol 12:592 (1998) (II:30) melanocortin1R (MSHR) mouse L98P, yet decr ag affinity Lu et al. Mol Endocrinol 12:592 (1998) [postulate agonist-mimetic] Robbins et al. Cell 72:827 (1993) sombre mutation L98P =-=-=-=-=-=-=-=-=-= III:0 melanocortin1R (MSHR) mouse D115E,V,K Lu et al. Mol Endocrinol 12:592 (1998) III:3 rhodopsin bovine E113Q [salt bridge with VII:11 K296, below, in OFF state] Cohen et al. Biochem 32:6111 (1993) III:4 melanocortin1R (MSHR) mouse D119R,K (not V), yet decr ag affinity Lu et al. Mol Endocrinol 12:592 (1998) [postulate agonist-mimetic] Robinson et al. Neuron 9:719 (1992) III:6 adenosine A2BR human F84L,S constit act Li et al. J Pharmacol Exp Ther. 2006 Oct 31 [Epub ahead of print] III:7 alpha1bAR hamster D125K>A increase in basal IP (PLC act) [postulate salt bridge with VII:4 K331, below] Porter et al. JBC 271(45):28318 (1996) III:7 delta OR mouse basal GTPgS binding D128A 261% but decr ag aff D128N 215% Befort et al. JBC 274(26):18574 (1999) D128H,K also constit act; D128R not expressed Cavalli et al. Neuroscience 93:1025 (1999) III:7 CCK2R (=CCKBR) human M134A incr basal IP's Beinborn et al. Mol Pharm 65:753 (2004) III:7 FSHR human T449A,I Smit et al. Annu Rev Pharmacol Toxicol 47:22.1 (2007) III:8 melanocortin1R (MSHR) mouse C123R,K (not A,E), yet decr ag affinity MC1R fox C125R Lu et al. Mol Endocrinol 12:592 (1998) [postulate agonist-mimetic] III:8 delta OR mouse basal GTPgS binding Y129A 205% Y129F 172% but decr ag aff Befort et al. JBC 274(26):18574 (1999) III:10 B2AR human C116F increase in basal Na+/H+ exchange but not cAMP (AC act) Zuscik et al. JBC 273(6):3401 (1998) III:10 alpha1bAR hamster C128F increase in basal IP (PLC act) but not PLA2 act Perez et al. Mol Pharm 49:112 (1996) C128W>T,M,Y,L>A,F basal IP's Porter et al. JBC 271(45):28318 (1996) III:10 AT1R rat N111G>A not I,F increase in basal IP; claim binds to agonist Noda et al. Biochem 35:16435 (1996) AT1aR rat N111S,A probably CA (basal IP3 not determined) [postulate interaction with N295 VII:14 in the OFF state, see below] Balmforth et al. JBC 272(7):4245 (1997) III:10 bradykinin B1R human N121A Leeb-Lundberg et al. JBC 276(12):8785 (2001) III:10 bradykinin B2R human short splice variant N113A Marie et al. Mol Pharm 55:92 (1999) III:10 PAFR guinea pig N100A incr basal arach release, IP's, AC inhib Ishii et al. JBC 272(12):7846 (1997) III:10 ORL1 human N133W incr basal G14, G16 and Gi/o signaling Kam et al. J Neurochem 83(6):1461 (2002) III:10 CXCR4 N119S,A not D,K constit act Zhang et al. JBC 277(25):24515 (2002) III:11 rhodopsin bovine (but not the opsin) G121X (X=W>I>L>V>T>S>A) dark act Han et al. JBC 272(37):23081 (1997) Han et al. PNAS USA 94:13442 (1997) III:11 TSHR human S505R Van Sande et al. J Clin Endo Metab 80(9):2577 (1995) incr basal cAMP not IPs Tonacchera et al. J Clin Endo Metb 81:547 (1996) III:11 CB1R cannabinoid R human F200A incr constit act (Gi) Shen et al. Eur J Pharmacol Jan 23, 2006 [epub] III:11 CCKBR human V138A incr basal IP's Blaker et al. Mol Pharm 58:399 (2000) III:11 H1R human S->C,T incr constit act. Jongejan et al. Nature Chem Biol 1:98 (2005) III:14 KSHV-GPCR D132A but not N further increases basal IP's (wt constit act) Ho et al. JBC 276(2):1376 (2001) III:15 TSHR human V509A Van Sande et al. J Clin Endo Metab 80(9):2577 (1995) III:18 LHR human L457R -> FMPP incr basal cAMP Latronico et al. J Clin Endo Metab 83:2435 (1998) LHR human L457R,K>H not A,D incr basal cAMP; incr basal IPs not evident except R>K when using more sensitive reporter gene assay Shinozaki et al. Mol Endo 15:972 (2001) III:18 FSHR human L460R but not K,A,D incr basal cAMP (all less FSH-responsive) Tao et al. 14:1272 (2000) III:18 B2AR human L124R,K,A incr basal cAMP; D poorly expressed Tao et al. 14:1272 (2000) III:18 m1R rat L116A Lu and Hulme, JBC 274(11):7309 (1999) L116H not constit act; Emax ~wt Lu and Hulme, JBC 275(8):5682 (2000) III:18 CB1R cannabinoid R human L207A incr constit act (Gs) D'Antona et al. Brain Res 2006 Jul 28 (Epub ahead of print) III:21 cAR1 Dictyostelium L100H constit act Zhang et al. Mol Biol Cell 16:562 (2005) III:21 CB1R cannabinoid R human T210I not A constit act D'Antona et al. Biochemistry 45(17):5606 (2006) III:22 m1R rat S120A Lu and Hulme, JBC 274(11):7309 (1999) S120H not constit act; Emax ~50% wt Lu and Hulme, JBC 275(8):5682 (2000) III:24 opsin (rhodopsin apoprotein) E134Q not D constit act Cohen et al. Biochem 32:6111 (1993) E134S,I,Q not F,L constit act Acharya et al. JBC 271(41):25406 (1996) III:24 prostanoid receptor FPR human E132T not D constit act Pathe-Neuschafer-Rube et al. Biochem J (Epub 2005 Jan 14) III:24 alpha1bAR incr basal IP D142T>I>V,Q,M>L,C,A,S,K>H,G,Y,P>F,R,W E,N ~wt Scheer et al. PNAS USA 94:808 (1997) III:24 B2AR D130N>A incr basal cAMP; also decr stability Rasmussen et al. Mol Pharm 56:175 (1999) III:24 CCK2R (=CCKBR) human E151A incr basal IP's Foucaud et al. Mol Pharmacol epub Nov 17 (2005) III:24 CXCR2 human D138V (analogous to KSHV GPCR) but not Q incr basal IP Burger et al. J Immunol 163:2017 (1999) III:24 histamine H2R rat D115A,N incr basal cAMP; also decr stability Alewijnse et al. Mol Pharm 57:890 (2000) III:24 angiotensin II AT1AR rat D125A incr basal IP Gaborik et al. Endocrinology 144:2220 (2003) III:24 vasopressin V2R human D136A incr basal cAMP; AVP affinity ~wt; decr expr Morin et al. FEBS Lett 441:470 (1998) III:24 muOR rat D164H,Q,Y,M not E incr basal act; decr stability Li et al. Biochem 40:12039 (2001) III:24 KSHV-GPCR V142D but not N further increases basal IP's (wt constit act) Ho et al. JBC 276(2):1376 (2001) III:25 alpha1bAR hamster R143K>H,D (H,D,A,N impaired ag stim; I,E no coupling) Scheer et al. Mol Pharm 57:219 (2000) III:25 adenosine A3R human R108A,K constit act both Gq (PLC) and Gi (AC inhib) Chen et al. BBRC 284:596 (2001) III:25 oxytocinR human R137A ~50% incr basal IP's Chini et al. Adv Exp Med Biol 449:367 (1998) Fanelli et al. Mol Pharm 56:214 (1999) III:25 cAR1 Dictyostelium I104N and other polar substitutions constit act Zhang et al. Mol Biol Cell 16:562 (2005) Zhang et al. Mol Microbiol 65:508 (2007) III:27 m5R human F130X (X=Q>C>M>S,E,H,K>R>G>T in terms of % constit act) Burstein et al. JBC 273(38):24322 (1998) III:31 m5R human R134X (X=D>N,E,T>Q>A>C>V>G in terms of % constit act) Burstein et al. JBC 273(38):24322 (1998) =-=-=-=-=-=-=-=-=-= IV:21 m1R rat A160G incr basal IP's Lu et al. JBC 276(36):34098 (2001) =-=-=-=-=-=-=-=-=-= V:3 alpha1bAR hamster A204V increase in basal IP (PLC act) and PLA2 act Hwa et al. JBC 271(14):7956 (1996) [postulate interacts with VI:23] V:4 m1R rat F190C not A, incr basal IP; also incr ACh potency/affinity Allman et al. Mol Pharm 58:175 (2000) V:7 CCKBR human L223A incr basal IP's Blaker et al. Mol Pharm 58:399 (2000) V:10 TRHR mouse F199A Colson et al. Mol Pharm 54:968 (1998) V:18 LHR human I542L -> FMPP increase in basal cAMP Laue et al. PNAS USA 92:1906 (1995) V:18 TSHR human V->L,F Smit et al. Annu Rev Pharmacol Toxicol 47:22.1 (2007) V:18 FSHR human I545T Smit et al. Annu Rev Pharmacol Toxicol 47:22.1 (2007) V:22 TSHR human Y->N Smit et al. Annu Rev Pharmacol Toxicol 47:22.1 (2007) V:29 m5R human T220V,C but not A,G,L,F Burstein et al. JBC 271(6):2882 (1996) =-=-=-=-=-=-=-=-=-= (VI:-5) D1AR F264I (the corresponding D1BR residue) CA like the wt D1BR Charpentier et al. JBC 271(45):28071 (1996) (VI:-5) CCK2R (=CCKBR) human L325E incr basal IP's Beinborn et al. J Biol Chem 273:14146 (1998) VI:-2 B2AR human E268A>Q incr basal cAMP; also decr stability Ballesteros et al. JBC 276(31):29171 (2001) VI:-2 m1R E360A increased basal IP production Hogger et al. JBC 270(13):7405 (1995) human m1R Kosugi et al. Mol Pharm 53:894 (1998) VI:-2 FSHR human D567G Simoni et al. Steroids 63:288 (1998) Haywood et al. Mol Endo 16:2582 (2002) D567G or del-D567 not found CA; FSHR harder to activate than LHR,TSHR? Schulz et al. Mol Endo 13:181 (1999) VI:-2 LHR human D564G -> FMPP found incr basal cAMP L>F>G>N>A>V not E; incr basal IP L not F,G,N,A,V,E Kosugi et al. Mol Pharm 53:894 (1998) D564K>deletion>G>N but not AAA insertion after 562 or 564 incr basal cAMP but not basal IP; Schulz et al. Mol Endo 13:181 (1999) VI:-2 TSHR human D619G Kosugi et al. FEBS Lett 356:291(1994) Parma et al. Nature 365:649(1993) del-D619 Wonerow et al. JBC 273(14):7900 (1998) VI:-1 alpha1bAR hamster K290H increase in basal IP Cotecchia et al. PNAS USA 87:2896 (1990) VI:2 alpha1aAR A271E>K incr basal IP Rossier et al. Mol Pharm 56:858 (1999) VI:2 alpha1bAR hamster A293E>K>R>C>F>I>Q>M>L>V>P>Y>W>H>T>G>D>N>S basal IP (PLC) Kjelsberg et al. JBC 267(3):1430 (1992) ...and PLA2 act, at least by A293E Perez et al. Mol Pharm 49:112 (1996) VI:2 alpha2aAR T348X constitutive activity (inhibition of dopaminergic AC stimulation) K>C>E>A>F>T (wt), increase in agonist affinity K>C,E>A>F this position numbered T373 in other papers and swissprot:a2aa_human Ren et al. JBC 268(22):16483 (1993) VI:2 B1AR human L322K > A,C,F incr basal cAMP not E,T,I (~wt) Lattion et al. FEBS Lett 457:302 (1999) VI:2 B2AR human (Cys-min background) L272C some incr basal cAMP Jensen et al JBC 276(12):9279 (2001) VI:2 D2R human short form T343R constit act Gi/o Wilson et al. J Neurochem 77:493 (2001) VI:2 5-HT1bR human mild CA w/ Galphao coexpr; T313R,Q>F,K,A,N; H not expr Pauwels et al. Biochem J 343:435 (1999) VI:2 5-HT2aR rat C322 R,K > E several measures (incr 5HT affinity, basal IP's) Egan et al. JPET 286:85 (1998) VI:2 5-HT2cR rat S312 K > F incr basal IP production, 5-HT affinity and potency Herrick-Davis J Neurochem 69:1138 (1997) VI:2 5-HT4R mouse A258L Claeysen et al. Mol Pharm 55:910 (1999) VI:2 adenosine A3R human A229E constit act both Gq (PLC) and Gi (AC inhib) Chen et al. BBRC 284:596 (2001) VI:2 LHR human A568V -> FMPP Latronico et al. J Clin Endo Metab 83:2435 (1998) VI:2 TSHR human A623I,V Kosugi et al. FEBS Lett 356:291 (1994) also A623S Russo et al. J Clin Endo Metab 80:1347 (1995) VI:2 PAFR human L231R constitutive IP production Parent et al. JBC 271(14):7949 (1996) VI:2 muOR rat T279K incr basal activity, not D (decr coupling) Huang et al. Biochem 40:13501 (2001) VI:2 CCK2R (=CCKBR) human V332E incr basal IP's Beinborn et al. Mol Pharm 65:753 (2004) VI:5 LHR human M571I -> FMPP incr basal cAMP but not IP production Kosugi et al. Hum Mol Genet 4:183 (1995) VI:5 TSHR human M626I Ringkananont et al. Mol Endocrinol epub Dec 8 2005 VI:6 LHR human A572V -> FMPP incr basal cAMP and IP production Yano et al. J Clin Endo Metab 80:1162 (1995) VI:8 opsin bovine M257X (X=Y>N>S>A,D,V,T,K,P,E,H,I,W,C,G,P,Q not L,R(not exp)) Han et al. Biochemistry 37(22):8253 (1998) VI:8 AT1AR rat I245T incr basal IP's Parnot et al. PNAS USA 97(13):7615 (2000) VI:8 m5R human I447S Spalding et al. JBC 273(34):21563 (1998) VI:8 TSHR human L629F incr basal cAMP but not IP's Parma et al. J Clin Endo Metab 82(8):2695 (1997) VI:8 melanocortin MC4R human L250Q,N,E>K not R,A,F incr basal act Proneth et al. Chem Biol Drug Des 67:215 (2006). VI:8 H1R human I420S,K,R,A,G,E not F constit act Bakker et al. Mol Pharm 2007 Oct 24; [Epub ahead of print] VI:9 LHR human I575L -> FMPP Laue et al. Biochem Mol Med 58:192 (1996) (not in UCSF library) VI:9 TSHR human I630L Parma et al. J Clin Endo Metab 82(8):2695 (1997) VI:10 LHR human F576I,G,Y not E incr basal cAMP but not IP production Yano et al. J Clin Endo Metab 82(8):2586 (1997) VI:10 TSHR human F631C,L incr basal cAMP but not IP production Kosugi et al. FEBS Lett 356:291 (1994) VI:11 m5R human A450V Spalding et al. JBC 273(34):21563 (1998) VI:11 LHR human T577I -> FMPP incr basal cAMP but not IP production Kosugi et al. Hum Mol Genet 4:183 (1995) VI:11 TSHR human T632I incr basal cAMP but not IP production Kosugi et al. FEBS Lett 356:291 (1994) T632A shown in Fig 1 of Neumann et al. Mol Endo 15:1294 (2001) T632A incr basal cAMP Urizar et al. JBC 280(17):17135 (2005) VI:12 opsin bovine F261V Han et al. JBC 271(50):32337 (1996) VI:12 alpha1bAR hamster F303L,V,W >A,C not Y,S,G,N,E incr basal IP's (L,V signif inc Emax; C sl incr Emax; W,A decr Emax; Y sl incr Emax; S decr Emax; G,N,E no coupling; all incr epinephrine affinity except Y) Chen et al. Biochem 41:588 (2002) VI:12 B2AR F282L>A>G incr basal cAMP not N (impaired) Chen et al. Biochem 41:6045 (2002) VI:12 m5R human F451X constit. act. X=V,T>L>C,A,N not Q,G,W,P Spalding et al. JBC 273(34):21563 (1998) VI:12 C5aR human F251A Whistler et al. Traffic 3(12):866 (2002) VI:12 LHR human D578G,Y -> FMPP found P dead (maybe not expressed), N like wt; incr basal cAMP Y,F>G,E,S,L; incr basal IP production Y,F,L but not G,E,S Kosugi et al. JBC 271(50):31813 (1996) D578Y,H incr basal cAMP Min and Ascoli Mol Endo 14:1797 (2000) VI:12 FSHR rat D580G (but not human D581G) constit act; human FSHR VI:7 M576T and VII:10 H615Y (the rat residues) allow D581G constit act Tao et al. Mol Endo 16:1881 (2002) VI:12 TSHR human D633Y,E incr basal cAMP but not IP production Kosugi et al. FEBS Lett 356:291 (1994) D633A not N incr expression-normalized basal cAMP Govaerts et al. JBC 276(25):22991 (2001) Urizar et al. JBC 280(17):17135 (2005) D633H,F,W incr basal cAMP and basal IPs, C incr basal cAMP; not K,R (impaired signaling), del (low surf expr) or N (~wt) Neumann et al. Mol Endo 15:1294 (2001) VI:12 LGR7 human splice var 1 D637Y, var 2 D603Y incr basal cAMP but not IPs Hsu et al. Mol Endo 14:1257 (2000) VI:15 LHR human C581R -> FMPP increase in basal cAMP Laue et al. PNAS USA 92:1906 (1995) VI:16 TRHR mouse W279A Colson et al. Mol Pharm 54:958 (1998) VI:16 bradykinin B2R human short splice variant W256F>Q Marie et al. Mol Pharm 55:92 (1999) VI:16 LHR human M->G Smit et al. Annu Rev Pharmacol Toxicol 47:22.1 (2007) VI:17 D1R human L286A (not Y, which has decr agonist affinity and efficacy; also, D2R human VI:17 L387A was found agonist-insensitive) Cho et al. Mol Pharm 50:1338 (1996) VI:18 alpha1BAR P309A "unpublished data" mentioned in Chen et al. EMBO J 19:4265 (2000) VI:18 TSHR human P639S incr basal cAMP Tonacchera et al. J Clin Endo Metab 83:492 (1998) VI:19 TRHR mouse Y282A,F slight constitutive activity Colson et al. Mol Pharm 54:958 (1998) VI:20 m5R human N459H,S constit. act. not Y,K,T,Q,C,D,A,G,V,F (may lig-bind) Spalding et al. JBC 273(34):21563 (1998) VI:20 m3R rat N507S slight CA; perhaps ligand-mimetic effect (A,D like wt) Bluml et al. JBC 269(22):18870 (1994) VI:23 alpha1aAR hamster M292L increase in basal IP (PLC act) and PLA2 act Hwa et al. JBC 271(14):7956 (1996) [postulate interacts with V:3] VI:26 m5R human S465X constit. act. X=K>R>F>V>C,L>A>W,P>E>G,D Spalding et al. Biochem 36:10109 (1997) VI:26 m1R human S388Y slight constit. act. at high expression levels Huang et al. Mol Pharm 56:775 (1999) VI:29 TSHR human N650Y Parma et al. J Clin Endo Metab 82(8):2695 (1997) incr basal cAMP not IPs Tonacchera et al. J Clin Endo Metb 81:547 (1996) =-=-=-=-=-=-=-=-=-= VII:-1 TSHR human V656F Fuhrer et al. J Clin Endo Metab 82:3885 (1997) VII:4 alpha1bR hamster K331E>A increase in basal IP [postulate salt bridge with III:7 D125, above] Porter et al. JBC 271(45):28318 (1996) VII:7 rhodopsin bovine A292E [postulate subst for Schiff base counterion] Rao et al. Nature 367:639 (1994) VII:8 m1R rat W405A incr basal IP's Lu et al. JBC 276(36):34098 (2001) VII:10 neurotensin NTS1R rat F358A incr basal IPs Barroso et al. Br J Pharmacol 135:997 (2002) VII:11 rhodopsin bovine K296X (X=E>G>A>T>C>Q>V>H>L>F not R) inverse corr size [salt bridge with III:3 E113, above, in OFF state] Cohen et al. Biochem 32:6111 (1993) VII:11 delta OR mouse Y308F incr basal GTPgammaS binding but decr ag aff Befort et al. JBC 274(26):18574 (1999) VII:13 TSHR human N670S Van Sande et al. J Clin Endo Metab 80(9):2577 (1995) incr basal cAMP not IPs Tonacchera et al. J Clin Endo Metb 81:547 (1996) VII:13 LHR rat N593R incr basal cAMP; K ~wt; A,Q,S decr function; D,E decr expr Angelova et al. Mol Endo 14:459 (2000) VII:13 H1R human N->C,S,L not Q incr constit act Jongejan et al. Nature Chem Biol 1:98 (2005) VII:14 AT1AR rat N295S increase in basal IP3 [postulate interaction with N111 III:10 in the OFF state; N111S,A may also be constit act] Balmforth et al. JBC 272(7):4245 (1997) However, N295D not CA Perlman et al. Mol Pharm 51:301 (1997) and N295A not CA Hunyady et al. Mol Pharm 54:427 (1998) VII:15 TSHR human C672Y Van Sande et al. J Clin Endo Metab 80(9):2577 (1995) VII:17 LHR rat N597Q incr basal cAMP; R decr function; A,D,K decr expr Angelova et al. Mol Endo 14:459 (2000) VII:17 TSHR human N674D not A incr expression-normalized basal cAMP Govaerts et al. JBC 276(25):22991 (2001) Urizar et al. JBC 280(17):17135 (2005) N674D incr basal cAMP and basal IP's, S incr basal cAMP; not A (impaired signaling) Neumann et al. Mol Endo 15:1294 (2001) VII:20 TSHR human L->V Smit et al. Annu Rev Pharmacol Toxicol 47:22.1 (2007) VII:21 5HT2CR human Y368A,C incr basal IP's; F no coupling; all incr ag aff Rosendorff et al. Molec Brain Res 84:90 (2000) 5HT2CR human Y368G,Q,N,M>S,C,H>T,V,P,L,A incr basal IP's; N "locked on" (unresponsive to inverse agonists); revertants suggest VII:21/VIII interaction Prioleau et al. JBC epub July 26 2002 VII:24 AT1AR rat L305Q incr basal IP's Parnot et al. PNAS USA 97(13):7615 (2000) ---------------------------------------------------------------------------- other CA mutations ---------------------------------------------------------------------------- III/IC2 5-HT2cR human wt vs. RNA-edited forms III:29,31,(33) constit activity INI (wt) ~ VSI,INV,VNV,ISV > VSV >> VGV basal IP's, EC50 shift, presence of high-affinity state; Niswender et al. JBC 274(14):9472 (1999) EC2 thrombinR human/Xenopus chimera (stretch of 8 amino acids from Xenopus) fewer Xenopus residues or all EC parts from Xenopus -> less CA Nanevicz et al. JBC 271(2):702 (1996) IC3 TSHR and LHR deletions, incr basal cAMP (but not IP's) TSHR del 613-621 V:30-32,G,VI:-4-0 ++ LHR del 558-566 + del 613-617 V:30-32,G,VI:-4 + LHR del 558-562 + del 618-621 VI:-3-0 + LHR del 563-566 + del 609-621 V:26-32,G,VI:-4-0 - del 613-624 V:30-32,G,VI:-4-3 - del 609-624 V:26-32,G,VI:-4-3 - del 609-617 V:26-32,G,VI:-4 ++ LHR del 554-562 ++ del 618-624 VI:-3-3 - del 615 V:32 - del 616 G - del 618 VI:-1 - del 619 VI:-2 ++ LHR del 564 ++ (ECM: so for ++, delete mainly the end of one TM but not much of the other, or delete VI:-2; need VI:1->) Wonerow et al. JBC 273(14):7900 (1998) TSHR; hypothesize important residues at IC end of VI are pulled down out of the membrane and are thus more accessible to Gsalpha Schulz et al. Mol Endo 13:181 (1999) LHR VI:-4-2 B2AR with 266-272 subst. by analogous alpha1bR hamster residues (L266S/K267R/H269K/L272A) increase in basal cAMP Samama et al. JBC 268(7):4625 (1993) VI:1,2 CB1R human L341A/A342L constit stim of Gs but not Gi (although both pathways are activated by agonist) Abadji et al. J Neurochem 72:2032 (1999) VI:1,2,5,6 m2R Gi/o coupling motif VTIL pushed toward cytoplasmic side by insertion of 1-4 alanines right after the L -> CA. However, analogous insertion in m3R abolished activity, and insertion of alanines before the motif VTIL (AALS in m3) yielded ~wt properties. Liu et al. JBC 271(11):6172 (1996) EC3 B2AR hamster replaced by analogous alpha1aAR rat sequence - postulate enhanced affinity for G protein rather than intrinsic R* tendency Zhao et al. Mol Pharm 53:524 (1998) EC3 TSHR human deletion 658-661 VII:(1,2),3,4 Parma et al. J Clin Endo Metab 82(8):2695 (1997) VIII beta 1AR human R384Q/E Delos Santos et al. JBC 2006 Feb 24 Epub truncation of Cterm tail - D1AR rat Chaar et al. J Neurochem 79:1047 (2001) truncation of Cterm tail - EP3 receptors Hasegawa et al. JBC 271(4):1857 (1996) truncation of Cterm tail - TRHR C335stop mild CA but decr TRH response Matus-Leibovitch et al. JBC 270(3):1041 (1995) truncation of Cterm tail - avian betaAR - CA in membranes but not when purified/reconstituted; postulate disruption of interaction with regulatory factors which decrease the activity of the full-length, wt receptor in the cell Parker and Ross JBC 266(15):9987 (1991) truncation of Cterm tail - 5-HT4R splice variants all apparently CA, but shorter splice variants are more so Claeysen et al. Mol Pharm 55:910 (1999) m2/m5 chimera Nterm->IC1 and TM6->Cterm from m2R Burstein et al. Biochem Pharmacol 51:539 (1996) ---------------------------------------------------------------------------- secretin family - Baldwin numbering resulting from Lichtarge alignment ---------------------------------------------------------------------------- II:7 H223R PTHR human Janssen-type chondrodysplasia Schipani et al. Science 258:98 (1995) also opossum PTHR but NOT rat PTHR, PTH2R, secretinR, calcitoninR, GHRHR, GLP1R, or CRHR H223R>K incr basal cAMP, other subst not CA (most decrease expression) Schipani et al. Mol Endo 11:851 (1997) H169R NOT CA in glucose-dependent insulinotropic peptide R (GIPR) rat Tseng and Lin Biochem Biophys Res Comm 232:96 (1997) II:7 H220R PTH1R Turner et al. JBC 273(7):3830 (1998) II:7 H180R PTH2R Turner et al. JBC 273(7):3830 (1998) Turner et al. Mol Endo 10:132 (1996) but note SecR H178D DECR AC stim II:7 H156R SecR rat slightly CA Ganguli et al. JPET 286:593 (1998) II:7 H178R glucagonR but not K,A,E CA in PTHR, glucagonR, VIP1R but not GLP1R, GIPR, calcitoninR Hjorth et al. Mol Endo 12:78 (1998) II:7 H178R but not K,A,D VPAC1R human (VIP1R) incr basal cAMP =strangely= mutations at known NTD VIP-binding residues (D68A or E36A) abolished CA although mutants were surface-expressed Gaudin et al. JBC 273(9):4990 (1998) =-=-=-=-=-=-=-=-=-= IV:-1 E261del,Q,A PAC1R rat incr basal cAMP; agonist affinity ~wt Cao et al. FEBS Lett 469:142 (2000) =-=-=-=-=-=-=-=-=-= VI:5 T410P PTHR human Janssen-type chondrodysplasia Schipani et al. NEJM 335:708 (1996) T410C,I>G,P,K,M,V,Q,A,L>R>E,N,H,D,S,W,F>Y~T(wt) basal cAMP Schipani et al. Mol Endo 11:851 (1997) VI:5 T322P SecR rat slightly CA Ganguli et al. JPET 286:593 (1998) VI:5 T352A glucagonR slightly CA but not P,S Hjorth et al. Mol Endo 12:78 (1998) VI:5 T2343P,K,A but not C,E,F VPAC1R human (VIP1R) incr basal cAMP =strangely= mutations at known NTD VIP-binding residues (D68A or E36A) abolished CA although mutants were surface-expressed Gaudin et al. Biochem Biophys Res Comm 254:15 (1999) VI:5 T340P GIPR rat incr basal cAMP Tseng and Lin Biochem Biophys Res Comm 232:96 (1997) =-=-=-=-=-=-=-=-=-= VII:20 PTHR I458R Schipani et al. J Clin Endo Metab 84:3052 (1999) ---------------------------------------------------------------------------- family 3 - Ca++-sensing receptors, metabotropic Glu receptors, GHRHR (authors' assignments of location taken) ---------------------------------------------------------------------------- Zhao et al. FEBS Lett 448:180 (1999) CaR human A843E TM7 but not K,V constit act; also CA with NTD deleted; A843E,K,V and A840E decrease Emax note previously reported mutants have incr Ca++ sensitivity, not true CA Jensen et al. JBC 275:29547 (2000) CaR human many mutants with multiple changes in the A116-P1136 region are CA (first half of NTD; random saturation mutagenesis) Mary et al. JBC 273(1):425 (1998) mGluR1 coupling to PLC in the absence of agonist inhibited by Ctail RRKK (877-890, "36 residues" after TM7) but longer-tail isoforms overcome this inhibition and are consitutively active ---------------------------------------------------------------------------- yeast mating factor receptors STE2 is the alpha-factor receptor, STE3 is the a-factor receptor ---------------------------------------------------------------------------- IC3 STE2 S. cerevisiae (231-243) -RSRRFLGLKQFDS- weak CA G237A, Q240A, F241A Clark et al. JBC 269(12):8831 (1994) IC3 STE3 S. cerevisiae L194Q,R>A but not F; various short IC3 deletions also CA Boone et al. 90:9921 (1993) III:? STE2 S. cerevisiae I142T, S145L, Q149R (mutated singly) Sommers et al. Biochemistry 39:6898 (2000) V:? STE2 S. cerevisiae A212V Lin et al. Biochemistry 44:1278 (2005) V:? STE2 S. cerevisiae S214C Dube et al. JBC 275(34):26492 (2000) S214C incr basal act 2x V:? STE2 S. cerevisiae L226W Sommers et al. Biochemistry 39:6898 (2000) VI:? STE2 S. cerevisiae Q253L Dube and Konopka Mol Cell Biol 18:7205 (1998) Sommers et al. Biochemistry 39:6898 (2000) Dube et al. JBC 275(34):26492 (2000) Q253C incr basal act 4.5x VI:? STE2 S. cerevisiae S254F Konopka et al. PNAS USA 93:6764 (1996) Dube and Konopka Mol Cell Biol 18:7205 (1998) F>G>L>W Dube et al. JBC 275(34):26492 (2000) S254C incr basal act 2x VI:? STE2 S. cerevisiae P258L Konopka et al. PNAS USA 93:6764 (1996) CD/NMR of wt and mutant peptides Arshava et al. Biopoly 46:343 (1998) Sommers et al. Biochemistry 39:6898 (2000) Dube et al. JBC 275(34):26492 (2000) P258C incr basal act 47x VI:? STE2 S. cerevisiae S259L Konopka et al. PNAS USA 93:6764 (1996) Abel et al. Biochim Biophys Acta 1448:12 (1998) S259P some conditions Sommers et al. Biochemistry 39:6898 (2000) S259P Dube et al. JBC 275(34):26492 (2000) S259C incr basal act 2x VI:? Q229P "towards extracellular end"; mating pheromone receptor Rcb2(6) of the fungus Coprinus cinereus Olesnicky et al. EMBO J 18:2756 (1999) VII:? STE2 S. cerevisiae A281T, L284H (mutated singly) Lin et al. Biochemistry 44:1278 (2005) VII:? STE2 S. cerevisiae S288A, S292A; these S may interact with Q253 Dube and Konopka Mol Cell Biol 18:7205 (1998) Sommers et al. Biochemistry 39:6898 (2000) S288P ---------------------------------------------------------------------------- SOME ISSUES.... some receptors just intrinsically easier to active than others; V-VI OFF-state interaction stronger in FSHR than in LHR Kudo et al. JBC 271(37):22470 (1996) ease of constitutive activation by mutation TSHR>LHR>FSHR Govaerts et al. JBC 276(25):22991 (2001) In similar expression system, rat delta but not mu opioid receptors signif constit act; constit act enhanced in the absence of Na+ Neilan et al. Brit J Pharmacol 128:556 (1999) many wt receptors have significant basal activity depending on expression stoichiometry relative to G proteins, level of expression of receptor and/or G protein, cell line, conditions, etc. Pauwels and Wurch, Mol Neurobiology 17(1-3):109 (1998) Table 1 Chen et al. Mol Pharm 57:125 (2000) Pauwels et al. JPET 292:654 (2000) pH Ghanouni et al. JBC 275(5):3121 (2000) constitutive activity of receptor can be conferred by coexpression of a G protein partner with low GDP affinity; example is coexpression of the long rather than the short splice variant of Galpha-s together with the B2AR Seifert et al. JBC 273:5109 (1998) counterexamples to activation by mutations at III:24: data for TXA2R alpha, citations for other receptors Capra et al. Mol Pharmacol (2004) epub Jun 30 ----------------------------------------------------------------------------