The partial primary structure of bovine rhodopsin and its topography in the retinal rod cell disc membrane.
The amino-terminal 39 amino acids of bovine rhodopsin have the sequence where both carbohydrate attachment sites (CHO) contain GlcNAc(3)Man(3). This region of rhodopsin's sequence is exposed at the internal membrane surface of the rod cell disc membrane. Rhodopsin's carboxyl-terminal 40 amino acids have the sequence where amino acid 1? is the carboxyl-terminal amino acid of rhodopsin. Serines and threonines in the sequence 6? ? 15? are phosphorylated by rhodopsin kinase in a light-dependent reaction. Trypsin can digest native rhodopsin, in the disc membrane at and thermolysin can hydrolyze bonds , and . Limited proteolysis by thermolysin at a site internal in the molecule has been exploited in order to prepare rhodopsin as two large fragments, F1 and F2. Cysteine(33)?, is highly reactive in the dark and is modified by N-ethylmaleimide and several alkylating agents. The carboxyl-terminal region 1?-39? reacts with the membrane-impermeable nitrene from N-(4-azido-2-nitrophenyl)-2-aminoethyl sulfonate and is therefore exposed at the external (cytoplasmic) surface of the disc membrane. 1-azldopyrene, a hydrophobic nitrene precursor, is being used to map those regions of the rhodopsin sequence which are located in a hydrophobic environment.
Hargrave
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School of Medicine, and Department of Chemistry and Biochemistry, Southern Illinois University at Carbondale, Carbondale, Illinois, USA 62901.
Fong
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Hugh McDowell
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Mas
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Curtis
D R
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Wang
J K
JK
Juszczak
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Smith
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