Effects of albumin and apolipoprotein C-II on the acyl-chain specificity of lipoprotein lipase catalysis.
In this study we used monoacid triacylglycerols of various acyl-chain lengths as substrates for probing the active-site structure and substrate specificity of lipoprotein lipase (LPL). An unexpected finding was that the albumin ligand binding site is accessible not only to long-chain fatty acids for its recognized functional role as a fatty acid acceptor, but also to short- and medium-chain monoacid triacylglycerol substrates. The observed striking inhibitory effect (99%) of albumin on the LPL-catalyzed hydrolysis of trihexanoylglycerol is probably the result of the high affinity interaction of albumin with this substrate. Spectrophotometric analyses indicated that there is one high affinity binding site per albumin molecule (apparent KD = 1.8 +/- 0.9 microM) for the interaction with trihexanoylglycerol. Despite LPL acyl-chain specificity being obscured by the substrate binding effect of albumin, a systematic study of the lipolysis reaction under various assay conditions demonstrated that tributyroylglycerol represents the best substrate for LPL, and the preferential order of LPL catalysis for both the basal and apoC-II-activated activities is: C4 > C6 > C8 > C10 > C12 > C18:1. In some assay conditions, the presence of albumin affects the above-mentioned order, which can be attributed to substrate binding by albumin, rather than an alteration in the specificity of LPL. The synergistic effect of apoC-II and albumin resulted in the preferential activation of LPL for the hydrolysis of long-chain triacylglycerols. Even with optimal assay conditions for the hydrolysis of long chain triacylglycerols, there is still a preferential reactivity of LPL with short- and medium-chain triacylglycerols.
Wang
C S
CS
Protein Studies Program, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Bass
H
H
Whitmer
R
R
McConathy
W J
WJ
D000818:Animals; D053304:Apolipoprotein C-II; D001056:Apolipoproteins C / Q000494:pharmacology*; D002384:Catalysis; D002417:Cattle; D002849:Chromatography, Gas; D007700:Kinetics; D008071:Lipoprotein Lipase / Q000378:metabolism*; D008892:Milk / Q000201:enzymology; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014280:Triglycerides / Q000737:chemistry / Q000378:metabolism
D053304:Apolipoprotein C-II; D001056:Apolipoproteins C; D014280:Triglycerides; D012710:Serum Albumin, Bovine; D008071:Lipoprotein Lipase