ADP-ribosylation of transducin by islet-activating protein. Identification of asparagine as the site of ADP-ribosylation

D. R. Manning, B. A. Fraser, R. A. Kahn, A. G. Gilman

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Islet-activating protein catalyzes the ADP-ribosylation of transducin, a guanine nucleotide-binding regulatory protein that mediates activation of a retinal cyclic GMP-selective phosphodiesterase. Radiolabel from [adenylate-32P]NAD+ was incorporated specifically into the α subunit of purified transducin. Maximal levels of incorporation approximated 0.8 mol of ADPribose/mol of transducin. A peptide containing the ADP-ribosyl moiety was purified from a tryptic digest of radiolableled transducin. This peptide was characterized by chemical and enzymatic procedures and by fast atom bombardment mass spectrometry. The primary structure of this peptide was Glu-Asn-Leu-Lys-Asn(ADP-ribose)-Gly-Leu-Phe. It is probable that the peptide originated from the carboxyl terminus of the α subunit and that the ADP-ribosyl moiety is attached by an N-glycosidic linkage to the asparagine residue. Transducin associated with retinal disc membranes is also ADP-ribosylated by cholera toxin. Cholera toxin and islet-activating protein sequentially catalyze the incorporation of 1.9 mol of ADP-ribose/mol of transducin, indicating two distinct sites of ADP-ribosylation within transducin.

Original languageEnglish (US)
Pages (from-to)749-756
Number of pages8
JournalJournal of Biological Chemistry
Volume259
Issue number2
StatePublished - 1984

Fingerprint

Transducin
Asparagine
Pertussis Toxin
Adenosine Diphosphate
Adenosine Diphosphate Ribose
Peptides
Cholera Toxin
glycylleucine
Fast Atom Bombardment Mass Spectrometry
Guanine Nucleotides
Cyclic GMP
Phosphoric Diester Hydrolases
GTP-Binding Proteins
NAD
Mass spectrometry
Carrier Proteins
Chemical activation
Membranes
Atoms

ASJC Scopus subject areas

  • Biochemistry

Cite this

ADP-ribosylation of transducin by islet-activating protein. Identification of asparagine as the site of ADP-ribosylation. / Manning, D. R.; Fraser, B. A.; Kahn, R. A.; Gilman, A. G.

In: Journal of Biological Chemistry, Vol. 259, No. 2, 1984, p. 749-756.

Research output: Contribution to journalArticle

Manning, D. R. ; Fraser, B. A. ; Kahn, R. A. ; Gilman, A. G. / ADP-ribosylation of transducin by islet-activating protein. Identification of asparagine as the site of ADP-ribosylation. In: Journal of Biological Chemistry. 1984 ; Vol. 259, No. 2. pp. 749-756.
@article{67e0215a48b9476f8163e4421e371d45,
title = "ADP-ribosylation of transducin by islet-activating protein. Identification of asparagine as the site of ADP-ribosylation",
abstract = "Islet-activating protein catalyzes the ADP-ribosylation of transducin, a guanine nucleotide-binding regulatory protein that mediates activation of a retinal cyclic GMP-selective phosphodiesterase. Radiolabel from [adenylate-32P]NAD+ was incorporated specifically into the α subunit of purified transducin. Maximal levels of incorporation approximated 0.8 mol of ADPribose/mol of transducin. A peptide containing the ADP-ribosyl moiety was purified from a tryptic digest of radiolableled transducin. This peptide was characterized by chemical and enzymatic procedures and by fast atom bombardment mass spectrometry. The primary structure of this peptide was Glu-Asn-Leu-Lys-Asn(ADP-ribose)-Gly-Leu-Phe. It is probable that the peptide originated from the carboxyl terminus of the α subunit and that the ADP-ribosyl moiety is attached by an N-glycosidic linkage to the asparagine residue. Transducin associated with retinal disc membranes is also ADP-ribosylated by cholera toxin. Cholera toxin and islet-activating protein sequentially catalyze the incorporation of 1.9 mol of ADP-ribose/mol of transducin, indicating two distinct sites of ADP-ribosylation within transducin.",
author = "Manning, {D. R.} and Fraser, {B. A.} and Kahn, {R. A.} and Gilman, {A. G.}",
year = "1984",
language = "English (US)",
volume = "259",
pages = "749--756",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "2",

}

TY - JOUR

T1 - ADP-ribosylation of transducin by islet-activating protein. Identification of asparagine as the site of ADP-ribosylation

AU - Manning, D. R.

AU - Fraser, B. A.

AU - Kahn, R. A.

AU - Gilman, A. G.

PY - 1984

Y1 - 1984

N2 - Islet-activating protein catalyzes the ADP-ribosylation of transducin, a guanine nucleotide-binding regulatory protein that mediates activation of a retinal cyclic GMP-selective phosphodiesterase. Radiolabel from [adenylate-32P]NAD+ was incorporated specifically into the α subunit of purified transducin. Maximal levels of incorporation approximated 0.8 mol of ADPribose/mol of transducin. A peptide containing the ADP-ribosyl moiety was purified from a tryptic digest of radiolableled transducin. This peptide was characterized by chemical and enzymatic procedures and by fast atom bombardment mass spectrometry. The primary structure of this peptide was Glu-Asn-Leu-Lys-Asn(ADP-ribose)-Gly-Leu-Phe. It is probable that the peptide originated from the carboxyl terminus of the α subunit and that the ADP-ribosyl moiety is attached by an N-glycosidic linkage to the asparagine residue. Transducin associated with retinal disc membranes is also ADP-ribosylated by cholera toxin. Cholera toxin and islet-activating protein sequentially catalyze the incorporation of 1.9 mol of ADP-ribose/mol of transducin, indicating two distinct sites of ADP-ribosylation within transducin.

AB - Islet-activating protein catalyzes the ADP-ribosylation of transducin, a guanine nucleotide-binding regulatory protein that mediates activation of a retinal cyclic GMP-selective phosphodiesterase. Radiolabel from [adenylate-32P]NAD+ was incorporated specifically into the α subunit of purified transducin. Maximal levels of incorporation approximated 0.8 mol of ADPribose/mol of transducin. A peptide containing the ADP-ribosyl moiety was purified from a tryptic digest of radiolableled transducin. This peptide was characterized by chemical and enzymatic procedures and by fast atom bombardment mass spectrometry. The primary structure of this peptide was Glu-Asn-Leu-Lys-Asn(ADP-ribose)-Gly-Leu-Phe. It is probable that the peptide originated from the carboxyl terminus of the α subunit and that the ADP-ribosyl moiety is attached by an N-glycosidic linkage to the asparagine residue. Transducin associated with retinal disc membranes is also ADP-ribosylated by cholera toxin. Cholera toxin and islet-activating protein sequentially catalyze the incorporation of 1.9 mol of ADP-ribose/mol of transducin, indicating two distinct sites of ADP-ribosylation within transducin.

UR - http://www.scopus.com/inward/record.url?scp=0021345742&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021345742&partnerID=8YFLogxK

M3 - Article

VL - 259

SP - 749

EP - 756

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 2

ER -