Purification strategies for Sendai virus membrane proteins.
J Chromatogr
; 397: 165-74, 1987 Jun 26.
Article
em En
| MEDLINE
| ID: mdl-2821034
ABSTRACT
Viral membrane proteins extracted from Sendai virions with the non-ionic detergents decylpolyethyleneglycol-300 and Triton X-100 were used as a model mixture of hydrophobic membrane proteins. The detergent extract contained the fusion protein (F) and the tetrameric and dimeric forms of the hemagglutinin-neuraminidase protein (HN). These proteins were purified by size-exclusion high-performance liquid chromatography (HPLC) in the presence of 0.1% sodium dodecyl sulphate, by ion-exchange and metal chelate affinity HPLC in the presence of 0.1% decylpolyethyleneglycol, and by reversed-phase HPLC without prior removal of the detergent. The tetramer of HN and F could be purified by size-exclusion HPLC after dissociation of a micellar aggregate containing tetrameric HN and multimeric F. The F and HN proteins could be purified by ion-exchange HPLC. Pure F protein could be obtained after metal chelate affinity HPLC. The F protein and the dimer and tetramer of HN could be eluted from a large-pore (100 nm) reversed-phase column, but they were eluted as broad, overlapping peaks. Only after reduction of the virion extract, the relatively small (13-15 kilodaltons) F2 protein could be obtained in pure form.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas da Matriz Viral
/
Vírus da Parainfluenza 1 Humana
Idioma:
En
Revista:
J Chromatogr
Ano de publicação:
1987
Tipo de documento:
Article
País de afiliação:
Holanda