Trypsin treatment may impair the interfacial activation action of lipoprotein lipase.
Lipoprotein lipase was expressed in Chinese hamster ovary (CHO) cells transfected with human lipoprotein lipase cDNA. The lipoprotein lipase retained tributyrin, water-soluble substrate, hydrolyzing activity (esterase activity). The catalytic action of this enzyme was studied by monitoring the esterase activity. The esterase activity was enhanced 4.5-fold by the addition of triolein emulsified with Triton X-100. This process was named interfacial activation. Treatment of LPL with trypsin (100 micrograms/ml, 37 degrees C for 10 min) caused the loss of the triolein hydrolyzing activity without that of the esterase activity. The esterase activity of trypsin-treated LPL was not enhanced by the addition of the triolein emulsion. The trypsin-treated LPL retained the ability to bind to very low density lipoproteins (VLDL). These results are consistent with the idea that LPL has a catalytic site and a lipid interface recognition site, and that the enzyme undergoes interfacial activation, in which the concealed catalytic site is revealed after the enzyme binds to the surface. Based on this hypothesis, the results obtained suggest that trypsin nicking may impair the interfacial activation process and cause the loss of the lipase activity.
Tashiro
J
J
Second Department of Internal Medicine, Chiba University.
Kobayashi
J
J
Shirai
K
K
Saito
Y
Y
Nakamura
H
H
Morimoto
Y
Y
Yoshida
S
S
D000818:Animals; D016466:CHO Cells / Q000201:enzymology / Q000502:physiology; D002850:Chromatography, Gel; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004655:Emulsions; D004789:Enzyme Activation; D004950:Esterases / Q000378:metabolism; D006801:Humans; D008071:Lipoprotein Lipase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D008079:Lipoproteins, VLDL / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D012685:Sepharose / Q000031:analogs & derivatives; D012967:Sodium Dodecyl Sulfate; D013462:Sulfur Radioisotopes; D013499:Surface Properties; D014162:Transfection; D014280:Triglycerides / Q000378:metabolism / Q000494:pharmacology; D014304:Triolein / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology*
D003470:Culture Media; D004655:Emulsions; D008079:Lipoproteins, VLDL; D013462:Sulfur Radioisotopes; D014280:Triglycerides; C037405:heparin-sepharose; D014304:Triolein; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; D004247:DNA; D012685:Sepharose; D008715:Methionine; D004950:Esterases; D008071:Lipoprotein Lipase; D014357:Trypsin; C005830:tributyrin