Nanoscale organization of multiple GPI-anchored proteins in living cell membranes

被引:673
作者
Sharma, P
Varma, R
Sarasij, RC
Ira
Gousset, K
Krishnamoorthy, G
Rao, M
Mayor, S
机构
[1] Natl Ctr Biol Sci TIFR, Bangalore 560065, Karnataka, India
[2] Raman Res Inst, Bangalore 560080, Karnataka, India
[3] Tata Inst Fundamental Res, Dept Chem Sci, Bombay 400005, Maharashtra, India
基金
英国惠康基金;
关键词
D O I
10.1016/S0092-8674(04)00167-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cholesterol and sphingolipid-enriched "rafts" have long been proposed as platforms for the sorting of specific membrane components including glycosylphosphatidylinositol-anchored proteins (GPI-APs), however, their existence and physical properties have been controversial. Here, we investigate the size of lipid-dependent organization of GPI-APs in live cells, using homo and hetero-FRET-based experiments combined with theoretical modeling. These studies reveal an unexpected organization wherein cell surface GPI-APs are present as monomers and a smaller fraction (20%-40%) as nanoscale (<5 nm) cholesterol-sensitive clusters. These clusters are composed of at most four molecules and accommodate diverse GPI-AP species; crosslinking GPI-APs segregates them from preexisting GPI-AP clusters and prevents endocytosis of the crosslinked species via a GPI-AP-selective pinocytic pathway. In conjunction with an analysis of the statistical distribution of the clusters, these observations suggest a mechanism for functional lipid-dependent clustering of GPI-APs.
引用
收藏
页码:577 / 589
页数:13
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