Reconstitution of yeast microsomal lipid flip‐flop using endogenous aminophospholipids

FEBS Letters

Mayinger, Peter²  Nicolson, Teresa¹ 

[1] Max-Planck-Institut für Entwicklungsbiologie, Spemannstrasse 35, D-72076 Tübingen, Germany;Zentrum für Molekulare Biologie, Universität Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany

关键词: Membrane biogenesis;    Phosphatidylethanolamine;    Phospholipid flip-flop;    Saccharomyces cerevisiae;    ER;    endoplasmic reticulum;    PtdCho;    phosphatidylcholine;    PtdEtn;    phosphatidylethanolamine;    PtdIns;    phosphatidylinositol;    PtdSer;    phosphatidylserine;    PtdOH;    phosphatidic acid;    TNBS;    2;    4;    6-trinitrobenzenesulfonic acid;    TNP-PtdEtn;    trinitrophenyl-derivative of PtdEtn;    FDNP-PtdEtn;    fluorodinitrophenyl-derivative of PtdEtn;   

DOI  :  10.1016/S0014-5793(00)01684-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The molecular basis of transbilayer movement or flipping of phospholipids in the endoplasmic reticulum is largely unknown. To circumvent the problems inherent to studies with artificial phospholipid analogs, we studied microsomal flip-flop of endogenous phosphatidylethanolamine in yeast. The transbilayer transport of phosphatidylethanolamine was measured in reconstituted proteoliposomes derived from microsomal detergent extracts. Our results demonstrate that flipping is protease sensitive but does not require metabolic energy. Our assay is the first to use the endogenous substrate of the so-called ‘flippase’ to study phospholipid translocation in endomembranes and may therefore be crucial for the understanding of the catalytic properties of this elusive enzyme.

【 授权许可】

Unknown