Point mutations in bovine opsin can be classified in four groups with respect to their effect on the biosynthetic pathway of opsin.
Expression in vitro with the recombinant baculovirus expression system showed correct biosynthesis and post-translational processing of "wild-type' bovine opsin with regard to translocation, glycosylation, palmitoylation and targeting. However, several of these processes were severely affected by point mutations. From the overall results of 16 mutants reported here, four groups were distinguished. One group significantly affected neither biosynthesis nor folding of opsin (D83N, P291A, A299C-V300A-P303G). A second group produced a truncated protein (R69H, Y301F), suggesting that these positions are essential for a correct translational process. A third group affected membrane translocation as well as glycosylation, which can be interpreted as interference with the function of a transfer signal. Substitutions at positions Glu-113, Glu-122, Glu-134, Arg-135 and Lys-248 belong to this category. A fourth group induced structural changes in the protein that led to heterogeneous distribution in the plasma membrane (E113Q/D, W265F, Y268S). Taking any functional consequences of these mutations into consideration, it seems that point mutations can have mosaic effects and therefore should be examined at several levels (folding, targeting, functional parameters).
DeCaluwé
G L
GL
Department of Biochemistry, University of Nijmegen, The Netherlands.
DeGrip
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D015870:Gene Expression / Q000235:genetics; D006031:Glycosylation; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D019308:Palmitic Acid / Q000378:metabolism; D017354:Point Mutation / Q000235:genetics*; D014176:Protein Biosynthesis / Q000235:genetics; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012243:Rhodopsin / Q000235:genetics / Q000378:metabolism; D017299:Rod Opsins / Q000096:biosynthesis / Q000235:genetics*; D018411:Spodoptera / Q000235:genetics; D013820:Thermolysin / Q000378:metabolism; D014162:Transfection / Q000235:genetics
D008565:Membrane Proteins; D011994:Recombinant Proteins; D017299:Rod Opsins; D019308:Palmitic Acid; D012243:Rhodopsin; D013820:Thermolysin