Ribonuclease-neutralized pancreatic microsomal membranes from livestock for in vitro co-translational protein translocation.
Anal Biochem
; 484: 102-4, 2015 Sep 01.
Article
en En
| MEDLINE
| ID: mdl-26050631
ABSTRACT
Here, we demonstrate that pancreatic microsomal membranes from pigs, sheep, or cattle destined for human consumption can be used as a valuable and ethically correct alternative to dog microsomes for cell-free protein translocation. By adding adequate ribonuclease (RNase) inhibitors to the membrane fraction, successful in vitro co-translational translocation of wild-type and chimeric pre-prolactin into the lumen of rough microsomes was obtained. In addition, the human type I integral membrane proteins CD4 and VCAM-1 were efficiently glycosylated in RNase-treated microsomes. Thus, RNase-neutralized pancreatic membrane fractions from pig, cow, or sheep are a cheap, easily accessible, and fulfilling alternative to canine microsomes.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Páncreas
/
Ribonucleasas
/
Porcinos
/
Ovinos
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Inhibidores Enzimáticos
/
Membranas Intracelulares
Aspecto:
Ethics
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Anal Biochem
Año:
2015
Tipo del documento:
Article