RECONSTITUTION OF LYSOSOMAL SULFATE TRANSPORT IN PROTEOLIPOSOMES

被引：6

作者：

KOETTERS, PJ ^{[1
]}

CHOU, HF ^{[1
]}

JONAS, AJ ^{[1
]}

机构：

[1] UNIV CALIF LOS ANGELES,LOS ANGELES CTY HARBOR MED CTR,DEPT PEDIAT,DIV MED GENET E4,TORRANCE,CA 90502

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 1995年 / 1244卷 / 2-3期

关键词：

LYSOSOME; SULFATE TRANSPORT; PROTEOLIPOSOME;

D O I：

10.1016/0304-4165(95)00036-B

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：

As part of a strategy to purify the lysosomal sulfate transporter, we developed a method for reconstitution of transport in artificial membrane vesicles. Lysosomal membranes were prepared from Percoll density gradient purified rat liver lysosomes and membrane proteins were solubilized using the non-ionic detergent, Triton X-100. The solubilized proteins were mixed with liposomes prepared by sonication of egg yolk lecithin and the detergent was removed by passage of the mixture over Bio-beads XAD2. The resulting proteoliposomes exhibited saturable sulfate transport with characteristics that were very similar to those observed in lysosomal membranes. Transport in proteoliposomes had a K-m of 155 mu M, exhibited pH dependence and was sensitive to inhibition by DIDS. Reconstitution of transport in proteoliposomes may be useful as an assay for purification of the lysosomal sulfate carrier.

引用

页码：311 / 316

页数：6

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