Identification of ATP citrate lyase as a phosphoprotein.

T. C. Linn, P. Srere

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Abstract

ATP citrate lyase has been purified from rat liver by a new procedure which results in high yields of an intact and stable enzyme. The pure lyase (specific activity approximately equal to 10 at 25 degrees C) exhibits a single protein band upon sodium dodecyl sulfate (SDS)-gel electrophoresis (Mr = 110,000). This procedure minimizes protease degradation that usually occurs when the enzyme is isolated by previously described isolation methods. In addition, the lyase is shown to be a phosphoprotein. 32P-labeled lyase has been purified from rat liver following an intraperitoneal injection of inorganic [32P]phosphate into the animals. It has been demonstrated that this phosphate (structural phosphate) behaves as a serine phosphate and is not the same as the enzyme-bound phosphate (catalytic phosphate) that is derived from ATP during the lyase reaction. There are 2 structural phosphate residues for each enzyme tetramer molecule. Removal of the structural phosphate has been accomplished using a partially purified phosphatase derived from rat liver. The dephospholyase has the same Vmax as the native phosphoenzyme. Evidence indicates that the structural phosphate resides in a protease-sensitive region of the native enzyme.

Original languageEnglish
Pages (from-to)1691-1698
Number of pages8
JournalJournal of Biological Chemistry
Volume254
Issue number5
Publication statusPublished - Mar 10 1979

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ATP Citrate (pro-S)-Lyase
Phosphoproteins
Phosphates
Lyases
Liver
Enzymes
Rats
Peptide Hydrolases
Phosphoserine
Intraperitoneal Injections
Electrophoresis
Phosphoric Monoester Hydrolases
Sodium Dodecyl Sulfate
Animals
Adenosine Triphosphate
Gels
Degradation
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of ATP citrate lyase as a phosphoprotein. / Linn, T. C.; Srere, P.

In: Journal of Biological Chemistry, Vol. 254, No. 5, 10.03.1979, p. 1691-1698.

Research output: Contribution to journalArticle

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