A model for studying LCAT reaction: In vitro cholesterol esterification in pig ovarian follicular fluid |
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Authors: | Jeffrey K Yao Steve C S Chang Robert J Ryan Peter James Dyck |
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Affiliation: | (1) Mayo Clinic and Mayo Foundation, 55901 Rochester, Minnesota |
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Abstract: | Phosphatidylcholine acyltransferase (lecithin:cholesterol acyltransferase or LCAT; EC 2.3.1.43) activity was found to be present
in pig ovarian follicular fluid (POFF), in addition to pig serum (PS). The cholesterol esterification rate in both POFF and
PS is linear with incubation time up to 2 hr. The mean absolute rate of POFF-cholesterol esterification was 8.1±0.4 nmoles
per ml per hr approximately one-fourth of that in PS. However, the fractional rate (percent of labeled cholesterol esterified
per hr) of POFF-cholesterol esterification was similar to that observed in PS. There was little variation of absolute rate
of cholesterol esterification in the fluid obtained from different sizes of follicles. Fatty acid or triacylglycerol did not
participate in the reaction of cholesterol esterification in POFF. No appreciable change in enzymatic activity was found from
storing POFF at 4 C for periods of time up to 24 hr or at −70 C up to 2 months, but activity was lost thereafter. On the other
hand, PS showed a much longer period of stability (5 days at 4 C and 9 months at −70 C). A discrepancy between the fatty acid
composition of cholesteryl esters formed by the LCAT reaction and the fatty acid composition at the C-2 position of phosphatidylcholine
led us to propose a two-step mechanism for the LCAT reaction. It is concluded that the LCAT of POFF, as well as that of plasma,
is specific for individual fatty acids rather than for the fatty acid composition of phosphatidylcholine. The fatty acid concentration
of lysophosphatidylcholine decreased during prolonged incubation times (6 to 21 hr) suggesting that the increased lysophosphatidylcholine
formed as a product of the LCAT reaction may be reused as substrate for the LCAT reaction or for hydrolysis by lysophosphatidylcholine
hydrolase.
Presented at the AOCS Meeting, New York, May 1977. |
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