In vivo phosphorylation of liver glycogen synthase. Isolation of the 32P-labeled enzyme and studies on the nature of the bound [32P]phosphates

A. W H Tan, frank q Nuttall

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Our previous study (Tan, A.W.H., and Nuttall, F.Q. (1983) J. Biol. Chem. 258, 9624-9630) indicated that liver synthase D contained a large number of endogenous phosphates, 12 of which were stable and 6 labile to alkali treatment. We wished to investigate the nature of the phosphates on synthase which became isotopically labeled when inorganic [32P]phosphate was given either to intact rats or to isolated liver cells. An antibody against liver synthase D was used for the isolation of synthase. The antibody recognized both the phosphorylated and dephosphorylated form of the enzyme, native as well as partially cleaved species. A large enzyme form, with M(r) of 90,000 as well as one with M(r) of 73,000 was observed. A 61% decrease in [32P]phosphate was found in synthase when prelabeled liver cells were treated with glucose, whereas a 25% increase was seen in cells treated with glucagon. After [32P]synthase D was converted to synthase I by synthase phosphatase, 95% of the [32P]phosphate was lost. All of the bound [32P]phosphates were found to be labile to alkali. Thus, under the in vivo conditions used, the [32P]phosphates incorporated into synthase were characterized by their fast turnover rate, alkali lability and susceptibility to the action of synthase phosphatase, both in vivo and in vitro. These criteria serve to distinguish them from the slower turning-over, alkali-stable phosphates found previously in both synthases D and I.

Original languageEnglish (US)
Pages (from-to)4751-4757
Number of pages7
JournalJournal of Biological Chemistry
Volume260
Issue number8
StatePublished - Jan 1 1985
Externally publishedYes

Fingerprint Dive into the research topics of 'In vivo phosphorylation of liver glycogen synthase. Isolation of the 32P-labeled enzyme and studies on the nature of the bound [32P]phosphates'. Together they form a unique fingerprint.

Cite this