title: Lipoprotein lipase: role of intramolecular disulfide bonds in enzyme catalysis.
abstract: Lipoprotein lipase (LPL) catalyzes the hydrolysis of the triacylglycerol component of triacylglycerol-rich lipoproteins. There are 4 cysteine pairs that are completely conserved among LPLs of all species known. We examined the functional importance of each of the cysteine pairs in enzyme catalysis by examining LPLs produced in Cos cells by transfection. Immunoreactive LPL was produced by vectors encoding the wildtype LPL and each of the 4 cysteine-pair mutant LPLs. Enzyme activity was detectable in the wildtype enzyme, but not in 3 of the 4 Cys-->Ser mutant enzymes (C216S/C239S, C264S/C275S, and C278S/C283S). Interestingly, LPL activity was also present in the mutant (C418S/C438S), which affects the C-terminal cysteine pair, with a specific activity approximately 50% higher than that of wildtype. There is evidence that LPL contains two distinct domains consisting of the N-terminal three-quarters of the sequence connected by a flexible region to the C-terminal domain comprising the rest of the molecule. The conservation of catalytic function despite the disruption of the only disulfide bridge in the C-terminal domain of LPL indicates that the two domains can function independently of each other in enzyme catalysis.
authors: Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
authors: J Y
authors: JY
authors: Lo
authors: L C
authors: LC
authors: Smith
authors: L
authors: L
authors: Chan
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D003545:Cysteine; D018076:DNA, Complementary; D004220:Disulfides; D006801:Humans; D008071:Lipoprotein Lipase / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D017354:Point Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D012694:Serine
chemical_list: D018076:DNA, Complementary; D004220:Disulfides; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D012694:Serine; D008071:Lipoprotein Lipase; D003545:Cysteine