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M9610509.TXT
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1996-01-30
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Document 0509
DOCN M9610509
TI Localization of the palmitoylation site in the transmembrane protein
p12E of Friend murine leukaemia virus.
DT 9601
AU Hensel J; Hintz M; Karas M; Linder D; Stahl B; Geyer R; Biochemisches
Institut am Klinikum der; Justus-Liebig-Universitat, Giessen, Germany.
SO Eur J Biochem. 1995 Sep 1;232(2):373-80. Unique Identifier : AIDSLINE
MED/96035869
AB Friend murine leukaemia virus complex was propagated on murine cells in
the presence of [9,10-3H]palmitic acid. Virus particles were harvested
from the culture supernatant and lysed with detergents. The viral
transmembrane protein, p12E, was isolated from the lysates by
size-exclusion chromatography and purified by narrowbore reverse-phase
HPLC. Analysis of the purified product by matrix-assisted laser
desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)
revealed that the protein is palmitoylated carrying one fatty acid
residue. The radiolabelled fatty acid was released by hydroxylamine
treatment at pH 7, indicating that acylation occurred via a thioester
linkage. For allocation of the acylation site, p12E was digested with
trypsin. The resulting peptides were either directly subjected to
MALDI-TOF-MS or fractionated by microbore reverse-phase HPLC prior to
mass spectrometry. The results revealed that p12E of Friend murine
leukaemia virus is acylated at a cysteine residue situated at the
C-terminal side of the putative transmembrane anchor of the polypeptide.
Fatty acid analysis of the purified acylpeptide demonstrated that p12E
carries almost exclusively palmitic acid.
DE Acylation Amino Acid Sequence Animal Binding Sites Fatty
Acids/CHEMISTRY Friend Virus/*CHEMISTRY/GENETICS/METABOLISM Gene
Products, env/*CHEMISTRY/GENETICS/METABOLISM Mice Molecular Sequence
Data Palmitic Acids/*CHEMISTRY/METABOLISM Peptide
Fragments/CHEMISTRY/GENETICS Protein Processing, Post-Translational
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Support, Non-U.S. Gov't JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).