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PMID: 10944123 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Initial enzyme for glycosylphosphatidylinositol biosynthesis requires PIG-P and is regulated by DPM2.

The EMBO journal ·Vol. 19 ·No. 16 ·2000-08-15 ·Pages 4402-11

Watanabe R, Murakami Y, Marmor MD, Inoue N, Maeda Y, Hino J, Kangawa K, Julius M, Kinoshita T

Abstract

Glycosylphosphatidylinositols (GPIs) are attached to the C-termini of many proteins, thereby acting as membrane anchors. Biosynthesis of GPI is initiated by GPI-N-acetylglucosaminyltransferase (GPI-GnT), which transfers N-acetylglucosamine from UDP- N-acetylglucosamine to phosphatidylinositol. GPI-GnT is a uniquely complex glycosyltransferase, consisting of at least four proteins, PIG-A, PIG-H, PIG-C and GPI1. Here, we report that GPI-GnT requires another component, termed PIG-P, and that DPM2, which regulates dolichol-phosphate-mannose synthase, also regulates GPI-GnT. PIG-P, a 134-amino acid protein having two hydrophobic domains, associates with PIG-A and GPI1. PIG-P is essential for GPI-GnT since a cell lacking PIG-P is GPI-anchor negative. DPM2, but not two other components of dolichol-phosphate-mannose synthase, associates with GPI-GnT through interactions with PIG-A, PIG-C and GPI1. Lec15 cell, a null mutant of DPM2, synthesizes early GPI intermediates, indicating that DPM2 is not essential for GPI-GnT; however, the enzyme activity is enhanced 3-fold in the presence of DPM2. These results reveal new essential and regulatory components of GPI-GnT and imply co-regulation of GPI-GnT and the dolichol-phosphate-mannose synthase that generates a mannosyl donor for GPI.

MeSH Terms
Amino Acid Sequence Animals CD59 Antigens/metabolism Carrier Proteins/genetics,metabolism Cell Line Cell Separation Cloning, Molecular DNA, Complementary/metabolism Dolichol Monophosphate Mannose/metabolism Expressed Sequence Tags Flow Cytometry Fluorescent Antibody Technique Fungal Proteins/chemistry Glycosylphosphatidylinositols/biosynthesis,genetics Hexosyltransferases Humans Mannosyltransferases Membrane Proteins/genetics,metabolism Mice Molecular Sequence Data N-Acetylglucosaminyltransferases/chemistry,metabolism Plant Proteins/chemistry Plasmids/metabolism Protein Binding Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Transfection
Chemicals
CD59 Antigens Carrier Proteins DNA, Complementary Dscr5 protein, mouse Fungal Proteins Glycosylphosphatidylinositols Membrane Proteins PIGH protein, human Plant Proteins phosphatidylinositol glycan-class A protein Dolichol Monophosphate Mannose DPM2 protein, human Hexosyltransferases Mannosyltransferases N-Acetylglucosaminyltransferases N-acetyllactosaminide beta-1,6-N-acetylglucosaminyltransferase PIGC protein, human PIGP protein, human
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Watanabe R
Department of Immunoregulation, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan.
Murakami Y
Marmor M D
Inoue N
Maeda Y
Hino J
Kangawa K
Julius M
Kinoshita T
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.16.4402
Published
2000-08-15
Pages
4402-11
Language
English
Region
England
NLM ID
8208664
PMCID
PMC302040
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/19.16.4402 PMC Full Text View journal details
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