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标题：Structure and phospholipid transfer activity of human PLTP: analysis by molecular modeling and site-directed mutagenesis
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作者：Jarkko Huuskonen ; Gerd Wohlfahrt ; Matti Jauhiainen 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：1999
卷号：40
期号：6
页码：1123-1130
语种：English
出版社：American Society for Biochemistry and Molecular Biology
摘要：The plasma phospholipid transfer protein (PLTP) is an important regulator of high density lipoprotein (HDL) metabolism. We have here, based on sequence alignments of the plasma LPS-binding/lipid transfer protein family and the X-ray structure of the bactericidal/permeability increasing protein (BPI), modeled the structure of PLTP. The model predicts a two-domain architecture with conserved lipid-binding pockets consisting of apolar residues in each domain. By site-directed mutagenesis of selected amino acid residues and transient expression of the protein variants in HeLa cells, the pockets are shown to be essential for PLTP-mediated phospholipid transfer. A solid phase ligand binding assay was used to determine the HDL-binding ability of the mutants. The results suggest that the observed decreases in phospholipid transfer activity of the N-terminal pocket mutants cannot be attributed to altered HDL-binding, but the C-terminal lipid-binding pocket may be involved in the association of PLTP with HDL. Further, the essential structural role of a disulfide bridge between cysteine residues 146 and 185 is demonstrated. The structural model and the mutants characterized here provide powerful tools for the detailed analysis of the mechanisms of PLTP function. —Huuskonen, J., G. Wohlfahrt, M. Jauhiainen, C. Ehnholm, O. Teleman, and V. M. Olkkonen. Structure and phospholipid transfer activity of human PLTP: analysis by molecular modeling and site-directed mutagenesis. J. Lipid Res. 1999. 40: 1123–1130.
关键词：HDL metabolism ; lipid transfer proteins ; homology modeling