Structural waters define a functional channel mediating activation of the GPCR, rhodopsin.
Structural water molecules may act as prosthetic groups indispensable for proper protein function. In the case of allosteric activation of G protein-coupled receptors (GPCRs), water likely imparts structural plasticity required for agonist-induced signal transmission. Inspection of structures of GPCR superfamily members reveals the presence of conserved embedded water molecules likely important to GPCR function. Coupling radiolytic hydroxyl radical labeling with rapid H(2)O(18) solvent mixing, we observed no exchange of these structural waters with bulk solvent in either ground state or for the Meta II or opsin states. However, the radiolysis approach permitted labeling of selected side chain residues within the transmembrane helices and revealed activation-induced changes in local structural constraints likely mediated by dynamics of both water and protein. These results suggest both a possible general mechanism for water-dependent communication in family A GPCRs based on structural conservation, and a strategy for probing membrane protein structure.
Angel
Thomas E
TE
Department of Pharmacology, Center for Proteomics and Bioinformatics, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4965, USA.
Gupta
Sayan
S
Jastrzebska
Beata
B
Palczewski
Krzysztof
K
Chance
Mark R
MR
D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002853:Chromatography, Liquid; D006868:Hydrolysis / Q000528:radiation effects; D017665:Hydroxyl Radical / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010103:Oxygen Isotopes; D010446:Peptide Fragments / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry*; D012243:Rhodopsin / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D014867:Water / Q000737:chemistry*; D014965:X-Rays
D010103:Oxygen Isotopes; D010446:Peptide Fragments; D043562:Receptors, G-Protein-Coupled; D012997:Solvents; D014867:Water; D017665:Hydroxyl Radical; D012243:Rhodopsin
Nature. 2008 Jul 10;454(7201):183-7
18563085
Chem Rev. 2007 Aug;107(8):3514-43
17683160
Structure. 2008 Aug 6;16(8):1157-65
18682217
J Biol Chem. 2003 Jan 31;278(5):3162-9
12427735
Annu Rev Biochem. 2006;75:743-67
16756510
Biophys Chem. 1995 Sep-Oct;56(1-2):79-87
7662872
Science. 1950 Feb 17;111(2877):179-81
15403120
Methods Enzymol. 2000;317:353-68
10829290
Science. 1998 Mar 20;279(5358):1940-3
9506944
Nature. 2008 Sep 25;455(7212):497-502
18818650
Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2290-5
12601165
Biochemistry. 1999 Jun 15;38(24):7617-23
10387000
Nature. 2006 Jan 5;439(7072):109-12
16280982
Methods Enzymol. 1982;81:240-6
6212740
J Biol Chem. 2003 Jul 4;278(27):24896-24903
12707280
Trends Pharmacol Sci. 2008 Feb;29(2):79-83
18194818
Annu Rev Biophys Biomol Struct. 2006;35:251-76
16689636
Proc Natl Acad Sci U S A. 2009 May 26;106(21):8555-60
19433801
Biochemistry. 2001 Jul 3;40(26):7761-72
11425302
J Biol Chem. 1995 Mar 10;270(10):5073-6
7890614
Proc Natl Acad Sci U S A. 2006 Aug 22;103(34):12729-34
16908857
Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):5982-7
11972040
Nature. 2007 Nov 15;450(7168):383-7
17952055
Rev Sci Instrum. 2008 Feb;79(2 Pt 1):025101
18315323
Angew Chem Int Ed Engl. 2006 Jun 26;45(26):4270-3
16586416
J Synchrotron Radiat. 2007 May;14(Pt 3):233-43
17435298
Pharmacol Rev. 2008 Mar;60(1):43-78
18321962
Nature. 2008 May 15;453(7193):363-7
18480818
Nat Struct Mol Biol. 2009 Feb;16(2):168-75
19182802
Biochemistry. 1993 Dec 28;32(51):14176-82
8260503
J Mol Biol. 2006 Mar 17;357(1):163-72
16414074
Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16123-8
17060607
J Biol Chem. 2000 Feb 25;275(8):5355-60
10681509
J Gen Physiol. 1963 Nov;47:215-40
14080814
J Mol Biol. 2005 Apr 8;347(4):803-12
15769471
Science. 2000 Aug 4;289(5480):739-45
10926528
Science. 2007 Nov 23;318(5854):1258-65
17962520
Proc Natl Acad Sci U S A. 2008 May 27;105(21):7439-44
18490656
Biochemistry. 1993 Sep 7;32(35):9165-71
8396426
Proc Natl Acad Sci U S A. 2007 Jan 30;104(5):1552-7
17251352
Science. 2007 Nov 23;318(5854):1266-73
17962519
J Mol Biol. 2004 Nov 5;343(5):1409-38
15491621
Chem Rev. 2008 Jan;108(1):74-108
18095715
J Mol Biol. 2004 Sep 10;342(2):571-83
15327956
Biochemistry. 2003 Mar 18;42(10):2759-67
12627940
Methods Enzymol. 1982;81:223-36
7098867
J Gen Physiol. 1953 Nov 20;37(2):189-200
13109155
Anal Chem. 2005 May 15;77(10):3029-37
15889890
Anal Chem. 1999 Sep 15;71(18):3965-73
10500483