The C-terminus of lipoprotein lipase is essential for biological function but contains no domain for glycosylphosphatidylinositol anchoring

T. Bruin, N. B. Groot, J. Jansen, J. J. Kastelein

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Abstract

In this study we present evidence that the C-terminus of lipoprotein lipase contains no glycosylphosphatidylinositol addition signal and is therefore not a glycosylphosphatidylinositol-anchored protein. Furthermore, we present additional evidence that the C-terminus of lipoprotein lipase is essential for biological function. Flow cytometric analysis and enzyme-activity monitoring experiments revealed no pool of lipoprotein lipase releasable by phosphatidylinositol-specific phospholipase present on the membrane of COS cells transfected with the human lipoprotein lipase gene while, in contrast, a heparin-releasable pool could be demonstrated. [14C]Ethanolamine, a constituent of the glycosylphosphatidylinositol anchor, was not incorporated into lipoprotein lipase during metabolic labeling. C-terminal deletion mutants were constructed and expressed in COS cells to investigate the presence of glycosylphosphatidylinositol addition signal on the C-terminus of human lipoprotein lipase (LPL). The specific activities of the mutants M442 [des-(Leu443-Gly448)-LPL] and M437 [des-(Cys438-Gly448)-LPL] were 78% and 59%, respectively, less than the wild type, while the M432 mutant [des-(Ala433-Gly449)-LPL] was catalytically inactive. Determination of the stability of the mutants revealed a decreased stability of the M437, compared with wild-type, whereas M442 showed the same stability. Flow cytometric analysis showed sustained membrane expression for all mutants including the inactive M432 mutant. These results suggest that the C-terminus of lipoprotein lipase is essential for maintaining intact catalytic activity but is not involved in any posttranslational proteolytic processing, including cleavage of a glycosylphosphatidylinositol addition signal. We therefore conclude that membrane-binding of the lipase is not mediated by such anchoring
Original languageEnglish
Pages (from-to)1019-1025
JournalEuropean Journal of Biochemistry / FEBS
Volume221
Issue number3
DOIs
Publication statusPublished - 1994

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