Pituitary protein lipolytic factor(s): Partial purification by isoelectric focusing (IEF)

E. De Palo, F. Ceriotti, C. De Palo, G. Federspil, C. Scandellari

Research output: Contribution to journalArticle

Abstract

Tests were made on highly purified bovine peptidal pituitary factors for lipolytic activity on rat adipose tissue. The lipolytic protein factor from bovine pituitary glands was isolated and characterized by ethanol precipitation, gel filtration, and isoelectric focusing. The supernatant of the homogenized tissue was precipitated with ethanol, the precipitate was lyophilized and resuspended in HCl 1 mmole/liter, the insoluble was discarded, and the supernatant lyophilized. A solution of the lyophilized fraction was fractionated by gel filtration on Sephadex G-75. The bands with lipolytic activity were pooled and lyophilized. Lipolytic activity was determined by incubation of epididimal rat adipose tissue and by successive measurements of free fatty acids and glycerol released in the incubation medium. The gel filtration with the highest lipolytic activity (20.4 μequiv/g per 4 hr and 17.0 μmole/g per 4 hr glycerol) was submitted to isoelectric focusing. The gel filtration still appeared highly heterogeneous, but most of the lipolytic activity was concentrated in the protein band at pH 8.6.

Original languageEnglish
Pages (from-to)455-468
Number of pages14
JournalJournal of Protein Chemistry
Volume2
Issue number6
DOIs
Publication statusPublished - Dec 1983

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Keywords

  • glycerol release
  • isoelectric focusing (IEF)
  • lipolysis
  • lipolytic activity
  • lipolytic hormones
  • pituitary lipid-mobilizing peptides

ASJC Scopus subject areas

  • Biochemistry

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