Expression, purification and characterization of recombinant human angiogenin in Pichia pastoris.
Biosci Biotechnol Biochem
; 76(7): 1384-8, 2012.
Article
em En
| MEDLINE
| ID: mdl-22785483
ABSTRACT
The potential of angiogenin (Ang) for clinical use has been highlighted in view of its important roles in inducing angiogenesis, facilitating cell proliferation, and inhibiting cell apoptosis. To produce soluble, correctly folded recombinant protein with a high yield, a DNA fragment encoding human Ang was inserted into eukaryotic expression vector pPIC9 and transformed into Pichia pastoris. The expression of recombinant human Ang (rhAng) accounted for about 70% of total secreted proteins. Purifying the Ang from the culture supernatant yielded 30 mg/L at 90% purity by chromatography with a SP Sepharose FF column. Biological assays indicated that rhAng can induce new blood-vessel formation, promote HeLa cell proliferation, increase Erk1/2 phosphorylation, and upregulate c-myc expression. Preparation of bioactive rhAng might lay the basis for further functional study, and might provide an effective strategy for large-scale production of soluble human Ang.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Pichia
/
Ribonuclease Pancreático
/
Proteínas Recombinantes
/
Expressão Gênica
/
Membrana Corioalantoide
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Biosci Biotechnol Biochem
Ano de publicação:
2012
Tipo de documento:
Article