Alkaline density gradient floatation of membranes:: Polypeptide composition of the mammalian peroxisomal membrane

被引：17

作者：

Gouveia, AMM

Reguenga, C

Oliveira, MEM

Eckerskorn, C

Sá-Miranda, C

Azevedo, JE

机构：

[1] Univ Porto, Inst Biol Mol & Celular, P-4150180 Porto, Portugal

[2] Univ Porto, Inst Ciencias Biomed Abel Salazar, P-4150180 Porto, Portugal

[3] Max Planck Inst Biochem, D-8000 Munich, Germany

[4] Inst Genet Med Jacinto Magalhaes, Porto, Portugal

来源：

ANALYTICAL BIOCHEMISTRY | 1999年 / 274卷 / 02期

关键词：

peroxisome; membrane proteins; alkaline extraction;

D O I：

10.1006/abio.1999.4281

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method for purification of the peroxisomal membrane from rat liver is described. The procedure consists of floating the (contaminated) peroxisomal membranes through an alkaline sucrose density gradient. A good resolution between the peroxisomal membrane and other membrane systems is achieved. Using these floated peroxisomal membranes we have determined that only 7.8 +/- 0.9% of the total peroxisomal protein is alkali resistant. The polypeptide composition of these highly pure peroxisomal membranes was analyzed by SDS-PAGE. Bands corresponding to polypeptides with apparent molecular masses of 15, 18, 22, 24, 26, 29, 35, 36, 38, 40, 52, 55, 70, 74-77, and 88 kDa are detected upon Coomassie blue staining of polyacrylamide gels. The identity of several of these polypeptides was determined by N-terminal sequencing and Western blotting analysis. (C) 1999 Academic Press.

引用

页码：270 / 277

页数：8

共 33 条

[1] Pex14p, a peroxisomal membrane protein binding both receptors of the two PTS-dependent import pathways [J].