The phosphorylation of choline acetyltransferase

Gordon Bruce, Louis B. Hersh

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Human placental Choline Acetyltransferase (ChAT) has been shown to be phosphorylated in vitro by kinases present in rat brain. Phosphorylation occurs at a single site with the exclusive phosphoamino acid being serine. ChAT phosphorylation was shown to be calcium, and not cyclic nucleotide, dependent and was inhibited by inhibitors of calcium/calmodulin protein kinases including anti-calmodulin anti-sera. ChAT phosphorylation was stimulated by calmodulin (9 fold) and, to a lesser extent, by phosphatidylserine (4 fold). These results indicate the involvement of a calcium/calmodulin and possibly also a calcium/phosopholipid kinase. This finding was confirmed by demonstrating ChAT phosphorylation using both purified multifunctional calcium/calmodulin protein kinase (CaMK) and calcium/phospholipid protein kinase C (PKC) from rat brain. A stoichiometric incorporation of 0.9 mol phosphate/mol ChAT was achieved by CaMK. Phosphorylated ChAT could be isolated from freshly prepared rat brain synaptosomes. The results obtained with this model system support the hypothesis that in vivo a fraction of ChAT exists phosphorylated.

Original languageEnglish (US)
Pages (from-to)613-620
Number of pages8
JournalNeurochemical Research
Volume14
Issue number7
DOIs
StatePublished - Jul 1989

Fingerprint

Phosphorylation
Choline O-Acetyltransferase
Calmodulin
Calcium
Calcium-Calmodulin-Dependent Protein Kinases
Protein Kinases
Rats
Brain
Phosphotransferases
Phosphoamino Acids
Synaptosomes
Cyclic Nucleotides
Phosphatidylserines
Serine
Protein Kinase C
Phospholipids
Phosphates
Serum

Keywords

  • Choline acetyltransferase
  • phosphorylation
  • protein kinase
  • regulation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

The phosphorylation of choline acetyltransferase. / Bruce, Gordon; Hersh, Louis B.

In: Neurochemical Research, Vol. 14, No. 7, 07.1989, p. 613-620.

Research output: Contribution to journalArticle

Bruce, Gordon ; Hersh, Louis B. / The phosphorylation of choline acetyltransferase. In: Neurochemical Research. 1989 ; Vol. 14, No. 7. pp. 613-620.
@article{cd5e1250cd0b4ca08c389b6840033079,
title = "The phosphorylation of choline acetyltransferase",
abstract = "Human placental Choline Acetyltransferase (ChAT) has been shown to be phosphorylated in vitro by kinases present in rat brain. Phosphorylation occurs at a single site with the exclusive phosphoamino acid being serine. ChAT phosphorylation was shown to be calcium, and not cyclic nucleotide, dependent and was inhibited by inhibitors of calcium/calmodulin protein kinases including anti-calmodulin anti-sera. ChAT phosphorylation was stimulated by calmodulin (9 fold) and, to a lesser extent, by phosphatidylserine (4 fold). These results indicate the involvement of a calcium/calmodulin and possibly also a calcium/phosopholipid kinase. This finding was confirmed by demonstrating ChAT phosphorylation using both purified multifunctional calcium/calmodulin protein kinase (CaMK) and calcium/phospholipid protein kinase C (PKC) from rat brain. A stoichiometric incorporation of 0.9 mol phosphate/mol ChAT was achieved by CaMK. Phosphorylated ChAT could be isolated from freshly prepared rat brain synaptosomes. The results obtained with this model system support the hypothesis that in vivo a fraction of ChAT exists phosphorylated.",
keywords = "Choline acetyltransferase, phosphorylation, protein kinase, regulation",
author = "Gordon Bruce and Hersh, {Louis B.}",
year = "1989",
month = "7",
doi = "10.1007/BF00964869",
language = "English (US)",
volume = "14",
pages = "613--620",
journal = "Neurochemical Research",
issn = "0364-3190",
publisher = "Springer New York",
number = "7",

}

TY - JOUR

T1 - The phosphorylation of choline acetyltransferase

AU - Bruce, Gordon

AU - Hersh, Louis B.

PY - 1989/7

Y1 - 1989/7

N2 - Human placental Choline Acetyltransferase (ChAT) has been shown to be phosphorylated in vitro by kinases present in rat brain. Phosphorylation occurs at a single site with the exclusive phosphoamino acid being serine. ChAT phosphorylation was shown to be calcium, and not cyclic nucleotide, dependent and was inhibited by inhibitors of calcium/calmodulin protein kinases including anti-calmodulin anti-sera. ChAT phosphorylation was stimulated by calmodulin (9 fold) and, to a lesser extent, by phosphatidylserine (4 fold). These results indicate the involvement of a calcium/calmodulin and possibly also a calcium/phosopholipid kinase. This finding was confirmed by demonstrating ChAT phosphorylation using both purified multifunctional calcium/calmodulin protein kinase (CaMK) and calcium/phospholipid protein kinase C (PKC) from rat brain. A stoichiometric incorporation of 0.9 mol phosphate/mol ChAT was achieved by CaMK. Phosphorylated ChAT could be isolated from freshly prepared rat brain synaptosomes. The results obtained with this model system support the hypothesis that in vivo a fraction of ChAT exists phosphorylated.

AB - Human placental Choline Acetyltransferase (ChAT) has been shown to be phosphorylated in vitro by kinases present in rat brain. Phosphorylation occurs at a single site with the exclusive phosphoamino acid being serine. ChAT phosphorylation was shown to be calcium, and not cyclic nucleotide, dependent and was inhibited by inhibitors of calcium/calmodulin protein kinases including anti-calmodulin anti-sera. ChAT phosphorylation was stimulated by calmodulin (9 fold) and, to a lesser extent, by phosphatidylserine (4 fold). These results indicate the involvement of a calcium/calmodulin and possibly also a calcium/phosopholipid kinase. This finding was confirmed by demonstrating ChAT phosphorylation using both purified multifunctional calcium/calmodulin protein kinase (CaMK) and calcium/phospholipid protein kinase C (PKC) from rat brain. A stoichiometric incorporation of 0.9 mol phosphate/mol ChAT was achieved by CaMK. Phosphorylated ChAT could be isolated from freshly prepared rat brain synaptosomes. The results obtained with this model system support the hypothesis that in vivo a fraction of ChAT exists phosphorylated.

KW - Choline acetyltransferase

KW - phosphorylation

KW - protein kinase

KW - regulation

UR - http://www.scopus.com/inward/record.url?scp=0024404459&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024404459&partnerID=8YFLogxK

U2 - 10.1007/BF00964869

DO - 10.1007/BF00964869

M3 - Article

C2 - 2506478

AN - SCOPUS:0024404459

VL - 14

SP - 613

EP - 620

JO - Neurochemical Research

JF - Neurochemical Research

SN - 0364-3190

IS - 7

ER -