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

• Type: Article in International Scientific Journal
• Date: 1999

Authors

gouveia, amm (Author) Other; reguenga, c (Author) Other; oliveira, mem (Author) Other; eckerskorn, c (Author) Other; sa-miranda, c (Author) Other; azevedo, je (Author) ICBAS

Journal

Title: Analytical Biochemistry

Vol. 274

Pages: 270-277

ISSN: 0003-2697

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 17 Citations

Scopus - 19 Citations

Other information

Authenticus ID: P-001-32S

DOI: 10.1006/abio.1999.4281

Abstract (EN): 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 8