RRC ID 42229
Author Yahiro K, Wada A, Yamasaki E, Nakayama M, Nishi Y, Hisatsune J, Morinaga N, Sap J, Noda M, Moss J, Hirayama T.
Title Essential domain of receptor tyrosine phosphatase beta (RPTPbeta) for interaction with Helicobacter pylori vacuolating cytotoxin.
Journal J Biol Chem
Abstract Helicobacter pylori produces a potent exotoxin, VacA, which causes progressive vacuolation as well as gastric injury. Although VacA was able to interact with two receptor-like protein tyrosine phosphatases, RPTPbeta and RPTPalpha, RPTPbeta was found to be responsible for gastric damage caused by VacA. To define the region of RPTPbeta involved in VacA binding, we made mutants of human cDNA RPTPbeta-B, a short receptor form of RPTPbeta. Immunoprecipitation experiments to assess VacA binding to RPTPbeta-B mutants indicated that five residues (QTTQP) at positions 747-751 of the extracellular domain of RPTPbeta-B (which is commonly retained in RPTPbeta-A, a long form of RPTPbeta) play a crucial role in its interaction with VacA, resulting in vacuolation as well as Git-1 phosphorylation. Transfected cells expressing deletion mutant Delta752, which lacks QTTQP, or the double point mutant Delta747 (T748A,T749A) had diminished vacuolation in response to VacA. Treatment of RPTPbeta-B and Delta747 (which have QTTQP at 747-751) with neuraminidase and O-glycosidase diminished their VacA binding, whereas chondroitinase ABC did not have an effect. No inhibitory effect of pleiotrophin, a natural RPTPbeta ligand, on VacA binding to RPTPbeta-B or Delta747 was observed, supporting the conclusion that the extracellular region of RPTPbeta-B responsible for VacA binding is different from that involved in binding pleiotrophin. These data define the region in the RPTPbeta extracellular domain critical for VacA binding, in particular the sequence QTTQP at positions 747-751 with crucial threonines at positions 748 and 749 and are consistent with a role for terminal sialic acids possibly because of threonine glycosylation.
Volume 279(49)
Pages 51013-21
Published 2004-12-3
DOI 10.1074/jbc.M406473200
PII S0021-9258(20)69388-7
PMID 15383529
MeSH Animals Bacterial Proteins / chemistry Bacterial Proteins / metabolism* Bacterial Proteins / physiology COS Cells Carrier Proteins / chemistry Cell Cycle Proteins / metabolism Cell Line Chondroitin ABC Lyase / chemistry Cricetinae Cytokines / chemistry DNA Glycosylases / metabolism DNA Primers / chemistry DNA, Complementary / metabolism Electrophoresis, Polyacrylamide Gel GTPase-Activating Proteins Gene Deletion Glycosylation Humans Immunoprecipitation Ligands Mice Mice, Knockout Models, Genetic Mutation Nerve Tissue Proteins / chemistry* Nerve Tissue Proteins / genetics Neuraminidase / metabolism Peptides / chemistry Phosphoproteins / metabolism Phosphorylation Point Mutation Polymerase Chain Reaction Protein Binding Protein Isoforms Protein Structure, Tertiary Protein Tyrosine Phosphatases / chemistry* Protein Tyrosine Phosphatases / genetics RNA / chemistry Receptor-Like Protein Tyrosine Phosphatases, Class 5 Reverse Transcriptase Polymerase Chain Reaction Sialic Acids / chemistry Signal Transduction Threonine / chemistry Transfection
IF 4.238
Times Cited 29
Human and Animal Cells BHK-21(RCB1423)