Hormone-sensitive lipase from bovine adipose tissue.
Biochim Biophys Acta
; 887(1): 51-7, 1986 Jun 16.
Article
em En
| MEDLINE
| ID: mdl-3708011
ABSTRACT
Hormone-sensitive lipase has been purified to near homogeneity from bovine perirenal adipose tissue. The purification method involves isoelectric precipitation at pH 5.0, followed by partial solubilisation in Triton N-101 and ion-exchange chromatography on DE-52. After additional solubilisation, the enzyme is further purified by chromatography on phenyl-Sepharose and heparin-Sepharose. This procedure can be completed within three working days and yields approx. 30 units of enzyme with a specific activity of 30 U/mg. The enzyme has been identified as a polypeptide of Mr 84 000 by affinity labelling with [3H]diisopropyl fluorophosphate. This polypeptide comprises approx. 60-80% of the protein in the final preparation, as judged by scanning densitometry of SDS-polyacrylamide gels stained with silver or with Coomassie blue R. The polypeptide of Mr 84 000 serves as a substrate for cyclic AMP-dependent protein kinase, phosphorylation correlating with activation of the lipase. Polyclonal antibody to the lipase has been raised in a rabbit and shown to specifically cross-react with the Mr 84 000 subunit.
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Base de dados:
MEDLINE
Assunto principal:
Tecido Adiposo
/
Esterol Esterase
/
Hormônios
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Ano de publicação:
1986
Tipo de documento:
Article