Phosphorylation of the recombinant spliced variants of the alpha-sub-unit of the stimulatory guanine-nucleotide binding regulatory protein (Gs) by the catalytic sub-unit of protein kinase A

N J Pyne, M Freissmuth, S Pyne

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Abstract

Both GS alpha-1 and GS alpha-4 were phosphorylated by the purified catalytic sub-unit of protein kinase A. Phosphate incorporation into 220 pmol and 190 pmol of GS alpha-4 and GS alpha-1 after a 1 hour incubation with kinase was 14 pmol and 10 pmol, respectively. These low levels of phosphorylation are due to the thermal lability of purified recombinant GS alpha. However, the phosphorylation was inhibited by guanine nucleotides (GDP-beta-S, GppNHp and GTP) and is, therefore, a specific event. We suggest that, as for GS alpha phosphorylation by protein kinase C (Pyne et al., 1992), the guanine nucleotide-free form of GS alpha is the most likely substrate. Guanine-nucleotides reduce the lifetime and, therefore availability for phosphorylation, of guanine-nucleotide free GS alpha. GS alpha phosphorylation by protein kinase A in vitro provides preliminary evidence that a similar phosphorylation of GS alpha may be an important regulatory event in cells.
LanguageEnglish
Pages1081-1086
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume186
Issue number2
DOIs
Publication statusPublished - 1992

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Phosphorylation
Guanine Nucleotides
Cyclic AMP-Dependent Protein Kinases
GTP-Binding Proteins
Carrier Proteins
Proteins
Protein Kinase C-alpha
Guanosine Triphosphate
Protein Kinase C
Phosphotransferases
Hot Temperature
Phosphates
Availability
Substrates

Keywords

  • autoradiography
  • brain
  • electrophoresis, polyacrylamide gel
  • escherichia coli
  • GTP-binding proteins
  • genetic variation
  • guanosine diphosphate
  • guanosine triphosphate
  • guanylyl imidodiphosphate
  • kinetics
  • macromolecular substances
  • molecular weight
  • phosphorus radioisotopes
  • protein kinases
  • RNA precursors
  • RNA splicing
  • recombinant proteins
  • thionucleotides

Cite this

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title = "Phosphorylation of the recombinant spliced variants of the alpha-sub-unit of the stimulatory guanine-nucleotide binding regulatory protein (Gs) by the catalytic sub-unit of protein kinase A",
abstract = "Both GS alpha-1 and GS alpha-4 were phosphorylated by the purified catalytic sub-unit of protein kinase A. Phosphate incorporation into 220 pmol and 190 pmol of GS alpha-4 and GS alpha-1 after a 1 hour incubation with kinase was 14 pmol and 10 pmol, respectively. These low levels of phosphorylation are due to the thermal lability of purified recombinant GS alpha. However, the phosphorylation was inhibited by guanine nucleotides (GDP-beta-S, GppNHp and GTP) and is, therefore, a specific event. We suggest that, as for GS alpha phosphorylation by protein kinase C (Pyne et al., 1992), the guanine nucleotide-free form of GS alpha is the most likely substrate. Guanine-nucleotides reduce the lifetime and, therefore availability for phosphorylation, of guanine-nucleotide free GS alpha. GS alpha phosphorylation by protein kinase A in vitro provides preliminary evidence that a similar phosphorylation of GS alpha may be an important regulatory event in cells.",
keywords = "autoradiography, brain, electrophoresis, polyacrylamide gel, escherichia coli, GTP-binding proteins, genetic variation, guanosine diphosphate, guanosine triphosphate, guanylyl imidodiphosphate, kinetics, macromolecular substances, molecular weight, phosphorus radioisotopes, protein kinases, RNA precursors, RNA splicing, recombinant proteins , thionucleotides",
author = "Pyne, {N J} and M Freissmuth and S Pyne",
year = "1992",
doi = "10.1016/0006-291X(92)90857-H",
language = "English",
volume = "186",
pages = "1081--1086",
journal = "Biochemical and Biophysical Research Communications",
issn = "1090-2104",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Phosphorylation of the recombinant spliced variants of the alpha-sub-unit of the stimulatory guanine-nucleotide binding regulatory protein (Gs) by the catalytic sub-unit of protein kinase A

AU - Pyne, N J

AU - Freissmuth, M

AU - Pyne, S

PY - 1992

Y1 - 1992

N2 - Both GS alpha-1 and GS alpha-4 were phosphorylated by the purified catalytic sub-unit of protein kinase A. Phosphate incorporation into 220 pmol and 190 pmol of GS alpha-4 and GS alpha-1 after a 1 hour incubation with kinase was 14 pmol and 10 pmol, respectively. These low levels of phosphorylation are due to the thermal lability of purified recombinant GS alpha. However, the phosphorylation was inhibited by guanine nucleotides (GDP-beta-S, GppNHp and GTP) and is, therefore, a specific event. We suggest that, as for GS alpha phosphorylation by protein kinase C (Pyne et al., 1992), the guanine nucleotide-free form of GS alpha is the most likely substrate. Guanine-nucleotides reduce the lifetime and, therefore availability for phosphorylation, of guanine-nucleotide free GS alpha. GS alpha phosphorylation by protein kinase A in vitro provides preliminary evidence that a similar phosphorylation of GS alpha may be an important regulatory event in cells.

AB - Both GS alpha-1 and GS alpha-4 were phosphorylated by the purified catalytic sub-unit of protein kinase A. Phosphate incorporation into 220 pmol and 190 pmol of GS alpha-4 and GS alpha-1 after a 1 hour incubation with kinase was 14 pmol and 10 pmol, respectively. These low levels of phosphorylation are due to the thermal lability of purified recombinant GS alpha. However, the phosphorylation was inhibited by guanine nucleotides (GDP-beta-S, GppNHp and GTP) and is, therefore, a specific event. We suggest that, as for GS alpha phosphorylation by protein kinase C (Pyne et al., 1992), the guanine nucleotide-free form of GS alpha is the most likely substrate. Guanine-nucleotides reduce the lifetime and, therefore availability for phosphorylation, of guanine-nucleotide free GS alpha. GS alpha phosphorylation by protein kinase A in vitro provides preliminary evidence that a similar phosphorylation of GS alpha may be an important regulatory event in cells.

KW - autoradiography

KW - brain

KW - electrophoresis, polyacrylamide gel

KW - escherichia coli

KW - GTP-binding proteins

KW - genetic variation

KW - guanosine diphosphate

KW - guanosine triphosphate

KW - guanylyl imidodiphosphate

KW - kinetics

KW - macromolecular substances

KW - molecular weight

KW - phosphorus radioisotopes

KW - protein kinases

KW - RNA precursors

KW - RNA splicing

KW - recombinant proteins

KW - thionucleotides

U2 - 10.1016/0006-291X(92)90857-H

DO - 10.1016/0006-291X(92)90857-H

M3 - Article

VL - 186

SP - 1081

EP - 1086

JO - Biochemical and Biophysical Research Communications

T2 - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 1090-2104

IS - 2

ER -