Ubiquitin with a non-ATP-dependent slow intrinsic proteolytic activity: a mild and rapid purification procedure.
Indian J Biochem Biophys
;
1992 Jun; 29(3): 303-5
Artigo
em Inglês
| IMSEAR
| ID: sea-28597
ABSTRACT
Ubiquitin has been purified to homogeneity, through a dialysis membrane having a NMW cutoff of 12 kDa, by taking advantage of its non-dialysable nature under these conditions. The dialysate was continuously recycled through a CM-52 cation exchange column at pH 4.5. The adsorbed fraction was eluted selectively at pH 7.2. Ubiquitin (25 mg) was obtained from 500 ml of packed RBCs. On SDS PAGE, ubiquitin showed varying mobility depending on the time of boiling in SDS. With 2 min of boiling, the molecular weight seemed to be 10.5 kDa, whereas 10 min of boiling resulted in a molecular weight of 8.5 kDa. Ubiquitin showed a slow intrinsic proteolytic activity against SDS-denatured beta-galactosidase in the absence of ATP. For the first 4 hr, there was no detectable degradation, but degradation was nearly complete after 8 hr. These data are not in agreement with those of Freid et al. [Proc. Natl Acad. Sci, USA, 84 (1987), 3685] who have reported a proteolytic activity comparable to that of other proteolytic enzymes.
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Endopeptidases
/
Búfalos
/
Dados de Sequência Molecular
/
Cinética
/
Ubiquitinas
/
Trifosfato de Adenosina
/
Cromatografia Líquida de Alta Pressão
/
Sequência de Aminoácidos
/
Eletroforese em Gel de Poliacrilamida
/
Eritrócitos
Idioma:
Inglês
Revista:
Indian J Biochem Biophys
Ano de publicação:
1992
Tipo de documento:
Artigo
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