Structure and function in rhodopsin: mapping light-dependent changes in distance between residue 65 in helix TM1 and residues in the sequence 306-319 at the cytoplasmic end of helix TM7 and in helix H8.
Spin-labeled double mutants of rhodopsin were produced containing a reference nitroxide at position 65, at the cytoplasmic termination of helix TM1, and a second nitroxide in the sequence of residues 306-319, which includes the cytoplasmic termination of helix TM7 and nearly the entire surface helix H8. Magnetic dipole-dipole interactions between the spins are analyzed to provide interspin distance distributions in both the dark and photoactivated states of rhodopsin. The distributions, apparently resulting from the conformational flexibility of the side chains, are found to be consistent with the structural model of rhodopsin in the dark state derived from crystallography. Photoactivation of the receptor triggers an increase in distance between residues in TM7, but not those in H8, relative to the reference at position 65 in TM1. The simplest interpretation of the result is a movement of the cytoplasmic portion of TM7 away from TM1 by 2-4 A.
Altenbach
C
C
Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095-7008, USA.
Cai
K
K
Klein-Seetharaman
J
J
Khorana
H G
HG
Hubbell
W L
WL
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003545:Cysteine / Q000235:genetics; D003593:Cytoplasm / Q000737:chemistry* / Q000235:genetics; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D008027:Light; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics; D017433:Protein Structure, Secondary / Q000235:genetics; D012243:Rhodopsin / Q000737:chemistry* / Q000235:genetics; D013113:Spin Labels; D013329:Structure-Activity Relationship
D010446:Peptide Fragments; D013113:Spin Labels; D012243:Rhodopsin; D003545:Cysteine