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The Journal of Lipid Research, Vol. 39, 2021-2030, October 1998
Copyright © 1998 by Lipid Research, Inc.
Biosynthesis and secretion of human plasma phospholipid transfer protein
Jarkko Huuskonena,
Matti Jauhiainena,
Christian Ehnholma, and
Vesa M. Olkkonena
a Department of Biochemistry, National Public Health Institute, Mannerheimintie 166, 00300 Helsinki, Finland
Correspondence to:
Vesa M. Olkkonen.
Plasma phospholipid transfer protein (PLTP) plays a critical role in lipoprotein metabolism and reverse cholesterol transport. We have studied the biosynthesis and secretion of PLTP using a stably transfected inducible HeLa cell line. Pulse-chase analysis revealed that: i) the major secreted forms of PLTP carry complex N-glycans; ii) N-glycosylation is crucial for PLTP secretion; iii) Endo H- resistant forms of PLTP could not be enriched using a 20°C temperature block, indicating that the transport of PLTP from the endoplasmic reticulum to the Golgi apparatus is exceptionally sensitive to low temperatures; and iv) treatment of the PLTP-producing cells with the reducing agent dithiothreitol caused a reversible secretion arrest, suggesting a role of disulfide bonds in the correct folding of PLTP. Transient expression of C-terminally truncated PLTP variants in COS cells demonstrated that: i ) the 30 C-terminal amino acids are dispensable for PLTP secretion, whereas deletion of 3550 residues results in a complete absence of secretion; and ii) the deletion of 30 C-terminal amino acid residues almost completely abolished the phospholipid transfer activity of PLTP.
The present study describes for the first time the biosynthesis of phospholipid transfer protein and provides tools for detailed elucidation of the structurefunction relationships in the protein.Huuskonen, J., M. Jauhiainen, C. Ehnholm, and V. M. Olkkonen. Biosynthesis and secretion of human plasma phospholipid transfer protein. J. Lipid Res. 1998. 39: 20212030.
Supplementary key words:
HDL metabolism, lipid transfer, PLTP, protein secretion

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Copyright © 1998 by the American Society for Biochemistry and Molecular Biology.
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