Enzymolysis of high density lipoprotein with a combination of membrane-immobilized esterase and trypsin.
Apolipoprotein A-1 (apo A-1), a major component of high density lipoprotein (HDL), was efficiently digested by membrane-immobilized trypsin after HDL was treated with membrane-immobilized esterase. Compared to treatment with membrane-immobilized trypsin alone, the relative amounts of apo A-1 polypeptides, m/z 1723.78 and m/z 1568.82, increased by 2.7- and 3.9-fold, respectively, when HDL was treated with membrane-immobilized esterase and trypsin. Furthermore, the efficient digestion of apo A-1 by trypsin was inhibited when HDL was treated with membrane-immobilized esterase in the presence of an esterase inhibitor, 6,9-diamino-2-ethoxyacridine (acrinol). The data indicate that the lipid components of lipoproteins are released by membrane-immobilized esterase. This method can be used to investigate the structure and function of other apolipoproteins.
Graduate School of Science and Engineering (Science Section) and Venture Business Laboratory, Ehime University, Matsuyama 790-8577, Japan.
Takahiro
T
Sakikawa
Youji
Y
Shimazaki
D000818:Animals; D016632:Apolipoprotein A-I / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D004950:Esterases / Q000378:metabolism*; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D014357:Trypsin / Q000378:metabolism*
D016632:Apolipoprotein A-I; D004800:Enzymes, Immobilized; D008075:Lipoproteins, HDL; D004950:Esterases; D014357:Trypsin