Glycerol allows low-temperature phase separation of membrane proteins solubilized in Triton X-114: application to the purification of plant cytochromes P-450 and b5.
Anal Biochem
; 197(1): 125-31, 1991 Aug 15.
Article
em En
| MEDLINE
| ID: mdl-1952054
The potentiality of the Triton X-114 phase separation technique for the purification of proteins from plant microsomal membranes has been investigated. It was shown that glycerol significantly lowers the cloud point of Triton X-114 solutions in water and of Triton X-114 solubilizates from microsomal membranes. It was also established that solubilized membrane components decrease the temperature of Triton X-114 micellar aggregation. Solubilization of microsomal membrane using detergent to protein ratios lower than 3.5, however, resulted in complete inhibition of detergent phase separation. Phase partitioning of Triton X-114 microsomal solubilizates, performed at low temperature (4 degrees C), in the presence of glycerol, provided a very fast and efficient step for the purification of cytochromes P-450 and b5. Conditions allowing optimal recoveries of these cytochromes have been defined.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Citocromos b5
/
Sistema Enzimático do Citocromo P-450
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
1991
Tipo de documento:
Article
País de afiliação:
França