PMID- 10085243
OWN - NLM
STAT- MEDLINE
DCOM- 19990623
LR  - 20181113
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 339 ( Pt 1)
DP  - 1999 Apr 1
TI  - Mammalian PIG-L and its yeast homologue Gpi12p are
      N-acetylglucosaminylphosphatidylinositol de-N-acetylases essential in
      glycosylphosphatidylinositol biosynthesis.
PG  - 185-92
AB  - Glycosylphosphatidylinositol (GPI) is used as a membrane anchor by many
      eukaryotic cell-surface proteins. The second step of GPI biosynthesis is
      de-N-acetylation of N-acetylglucosaminylphosphatidylinositol (GlcNAc-PI). We have
      previously cloned the rat PIG-L gene by expression cloning that complemented a
      mutant Chinese hamster ovary cell line defective in this step. Here we show that 
      recombinant rat PIG-L protein purified from Escherichia coli as a complex with
      GroEL has GlcNAc-PI de-N-acetylase activity in vitro. The activity was not
      enhanced by GTP, which is known to enhance the de-N-acetylase activity of
      mammalian cell microsomes. As with other de-N-acetylases that act on the GlcNAc
      moiety, metal ions, in particular Mn2+ and Ni2+, enhanced the enzyme activity of 
      PIG-L. The Saccharomyces cerevisiae YMR281W open reading frame encodes a protein 
      (termed Gpi12p) with 24% amino acid identity with rat PIG-L. On transfection into
      mammalian PIG-L-deficient cells, this gene, GPI12, restored the cell-surface
      expression of GPI-anchored proteins and GlcNAc-PI de-N-acetylase activity. The
      disruption of the gene caused lethality in S. cerevisiae. These results indicate 
      that GlcNAc-PI de-N-acetylase is conserved between mammals and yeasts and that
      the de-N-acetylation step is also indispensable in yeasts.
FAU - Watanabe, R
AU  - Watanabe R
AD  - Department of Immunoregulation, Research Institute for Microbial Diseases, Osaka 
      University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan.
FAU - Ohishi, K
AU  - Ohishi K
FAU - Maeda, Y
AU  - Maeda Y
FAU - Nakamura, N
AU  - Nakamura N
FAU - Kinoshita, T
AU  - Kinoshita T
LA  - eng
SI  - GENBANK/AB017165
SI  - GENBANK/AB017166
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (DNA Primers)
RN  - 0 (Glycosylphosphatidylinositols)
RN  - 0 (Recombinant Proteins)
RN  - EC 3.5.- (Amidohydrolases)
RN  - EC 3.5.1.89 (N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase)
SB  - IM
MH  - Amidohydrolases/chemistry/genetics/*metabolism
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - CHO Cells
MH  - Cricetinae
MH  - DNA Primers
MH  - Genetic Complementation Test
MH  - Glycosylphosphatidylinositols/*biosynthesis
MH  - Molecular Sequence Data
MH  - Rats
MH  - Recombinant Proteins/chemistry/genetics/metabolism
MH  - Saccharomyces cerevisiae/*enzymology
MH  - Sequence Homology, Amino Acid
PMC - PMC1220143
EDAT- 1999/03/23 00:00
MHDA- 1999/03/23 00:01
CRDT- 1999/03/23 00:00
PHST- 1999/03/23 00:00 [pubmed]
PHST- 1999/03/23 00:01 [medline]
PHST- 1999/03/23 00:00 [entrez]
PST - ppublish
SO  - Biochem J. 1999 Apr 1;339 ( Pt 1):185-92.