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PMID: 2114095 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

An insulin-sensitive cytosolic protein kinase accounts for the regulation of ATP citrate-lyase phosphorylation.

The Biochemical journal ·Vol. 268 ·No. 3 ·1990-06-15 ·Pages 539-45

Yu KT, Benjamin WB, Ramakrishna S, Khalaf N, Czech MP

Abstract

Purified rat liver ATP citrate-lyase is phosphorylated on serine residues by an insulin-stimulated cytosolic kinase activity partially purified from rat adipocytes [Yu, Khalaf & Czech (1987) J. Biol. Chem. 262, 16677-16685]. The Km for lyase phosphorylation by this hormone-sensitive kinase activity is approx. 3 microM. Two-dimensional tryptic-peptide mapping of the 32P-labelled lyase reveals that the kinase-catalysed phosphorylation occurs primarily on a specific peptide. In intact 32P-labelled adipocytes, insulin enhances the serine phosphorylation of ATP citrate-lyase by 2-3-fold. Tryptic digestion of the 32P-labelled lyase immunopurified from insulin-treated adipocytes also yields one major phosphopeptide. 32P-labelled lyase tryptic peptides derived from labelling experiments in vitro and in vivo exhibit identical electrophoretic and chromatographic migration profiles. Furthermore, radio-sequencing of the phosphopeptide from lyase 32P-labelled in vitro indicates that serine-3 from the N-terminus is phosphorylated by the insulin-stimulated cytosolic kinase, in agreement with previous studies on the position of the phosphoserine residue in ATP citrate-lyase isolated from insulin-treated cells. Taken together, the similarity in site-specific phosphorylation of ATP citrate-lyase from insulin-treated adipocytes to that catalysed by the hormone-activated cytosolic kinase in vitro strongly suggests that this kinase mediates insulin action on lyase phosphorylation in intact cells.

MeSH Terms
ATP Citrate (pro-S)-Lyase/metabolism Adipose Tissue/cytology,enzymology Animals Epididymis/enzymology In Vitro Techniques Insulin/pharmacology Male Peptide Fragments/metabolism Peptide Mapping Phosphorylation Protein Kinases/metabolism Rats Rats, Inbred Strains Serine/metabolism
Chemicals
Insulin Peptide Fragments Serine ATP Citrate (pro-S)-Lyase Protein Kinases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Yu K T
Department of Physiology and Biophysics, School of Medicine, State University of New York, Stony Brook 11794.
Benjamin W B
Ramakrishna S
Khalaf N
Czech M P
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2680539
Published
1990-06-15
Pages
539-45
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1131472
Subset
IM
Grants
NIDDK NIH HHS · DK32520 · United States
NIDDK NIH HHS · DK39625 · United States
External Links
PubMed source DOI: 10.1042/bj2680539 PMC Full Text View journal details
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