Detection of glycosyl‐phosphatidylinositol‐anchored proteins on the surface of Nicotiana tabacum protoplasts

FEBS Letters

Dry, Ian B¹  Soole, Kathleen L²  Takos, Adam M² 

[1] CSIRO Division of Horticulture, Hartley Grove, Urrbrae, SA 5064, Australia;Centre for Plant Membrane Biology, School of Biological Sciences, The Flinders University of South Australia, GPO Box 2100, Adelaide, SA 5001, Australia

关键词: Plant;    GPI-anchored;    Biotin;    Plasma membrane;    GPI;    glycosyl-phosphatidylinositol;    TX-114;    octylphenoxypolyethoxyethanol;    PtdIns-PLC;    phosphatidylinositol-specific phospholipase C;    NHS-LC-biotin;    sulfosuccinimidyl-6-(biotinamido) hexanoate;   

DOI  :  10.1016/S0014-5793(97)00064-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Glycosyl-phosphatidylinositol (GPI)-anchored plasma membrane proteins have been found to be widespread in eukaryotes and protozoa but have not been reported in higher terrestrial plants. A sensitive biotin-based assay has been used to detect the presence of GPI-anchored proteins on the outer surface of cultured Nicotiana tabacum cells. Six proteins with molecular weights of 92, 84, 60.5, 54.5, 39.5 and 37 kDa were found to move from a Triton X-114 detergent-rich phase to an aqueous phase following incubation with phosphatidylinositol-specific phospholipase C (PtdIns-PLC). The behaviour of these proteins is consistent with the presence of a GPI-anchor. Seven GPI-anchored proteins were also detected on the surface of tobacco leaf protoplasts with molecular weights of 67.5, 62, 39, 33.5, 27, 23 and 15.6 kDa. These data demonstrate the presence of multiple GPI-anchored proteins on the plasma membrane of higher plant cells. © 1997 Federation of European Biochemical Societies.

【 授权许可】

Unknown