Medium-chain versus long-chain triacylglycerol emulsion hydrolysis by lipoprotein lipase and hepatic lipase: implications for the mechanisms of lipase action.
To explore how enzyme affinities and enzyme activities regulate hydrolysis of water-insoluble substrates, we compared hydrolysis of phospholipid-stabilized emulsions of medium-chain (MCT) versus long-chain triacylglycerols (LCT). Because substrate solubility at the emulsion surface might modulate rates of hydrolysis, the ability of egg yolk phosphatidylcholine to solubilize MCT was examined by NMR spectroscopy. Chemical shift measurements showed that 11 mol % of [13C]carbonyl enriched trioctanoin was incorporated into phospholipid vesicles as a surface component. Similar methods with [13C]triolein showed a maximum solubility in phospholipid bilayers of 3 mol % (Hamilton & Small, 1981). Line widths of trioctanoin surface peaks were half that of LCT, and relaxation times, T1, were also shorter for trioctanoin, showing greater mobility for MCT in phospholipid. In assessing the effects of these differences in solubility on lipolysis, we found that both purified bovine milk lipoprotein lipase and human hepatic lipase hydrolyzed MCT at rates at least 2-fold higher than for LCT. With increasing concentrations of MCT, saturation was not reached, indicating low affinities of lipase for MCT emulsions, but with LCT emulsion incubated with lipoprotein lipase, saturation was reached at relatively low concentration, demonstrating higher affinity of lipase for LCT emulsions. Differences in affinity were also demonstrated in mixed incubations where increasing amounts of LCT emulsion resulted in decreased hydrolysis of MCT emulsions. Increasing MCT emulsion amounts had little or no effect on LCT emulsion hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS)
Deckelbaum
R J
RJ
Department of Pediatrics, Columbia University, New York, New York 10032.
Hamilton
J A
JA
Moser
A
A
Bengtsson-Olivecrona
G
G
Butbul
E
E
Carpentier
Y A
YA
Gutman
A
A
Olivecrona
T
T
D000818:Animals; D053304:Apolipoprotein C-II; D001056:Apolipoproteins C / Q000302:isolation & purification; D001667:Binding, Competitive; D002417:Cattle; D004655:Emulsions; D006801:Humans; D006868:Hydrolysis; D008049:Lipase / Q000378:metabolism*; D008071:Lipoprotein Lipase / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D009682:Magnetic Resonance Spectroscopy; D012995:Solubility; D013379:Substrate Specificity; D014280:Triglycerides / Q000378:metabolism*
D053304:Apolipoprotein C-II; D001056:Apolipoproteins C; D004655:Emulsions; D014280:Triglycerides; D008049:Lipase; D008071:Lipoprotein Lipase