Cloning, overexpression, purification and preliminary characterization of human septin 8.
Protein J
; 29(5): 328-35, 2010 Jul.
Article
em En
| MEDLINE
| ID: mdl-20544379
ABSTRACT
Mammalian septins comprise a family of 14 genes that encode GTP-binding proteins involved in important cellular processes such as cytokinesis and exocytosis. Expression of three different constructs encoding human septin 8 were analyzed and the results show that SEPT8GC, a clone expressing the conserved domain plus C-terminal domain of human septin 8 yields the highest amount of recombinant protein. This protein was purified by affinity chromatography followed by a gel filtration chromatography. CD spectrum of SEPT8GC is characteristic of folded proteins and it presents a transition profile with a T (m) of 54 degrees C. Fluorescence emission spectra, analytic gel filtration and DLS reflect the sample oligomeric heterogeneity with the predominance of dimers in solution. Homology models indicate clearly that the preferred dimer interface is the one comprising the GTP binding site.
Texto completo:
1
Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Ligação ao GTP
/
Proteínas do Citoesqueleto
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Protein J
Ano de publicação:
2010
Tipo de documento:
Article