Active site labeling of a receptor-like protein tyrosine phosphatase
The inactivation of the cytoplasmic domain of rat LAR, a receptor-like protein tyrosine phosphatase (PTPase), by iodoacetate and not by iodoacetamide suggested that iodoacetate interacts in a highly selective manner with the enzyme. The data indicate that iodoacetate binds at the active site of the...
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| Veröffentlicht in: | The Journal of biological chemistry 1992-01, Vol.267 (1), p.140-143 |
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| creator | Pot, D A Dixon, J E |
| description | The inactivation of the cytoplasmic domain of rat LAR, a receptor-like protein tyrosine phosphatase (PTPase), by iodoacetate and not by iodoacetamide suggested that iodoacetate interacts in a highly selective manner with the enzyme. The data indicate that iodoacetate binds at the active site of the enzyme with a stoichiometry of 0.8 mol of iodoacetate bound per mol of rat LAR. A single [14C]iodoacetate-labeled peptide was isolated following endoproteinase Lys-C digestion of the radiolabeled PTPase. Sequence analysis of the active site labeled peptide demonstrates that Cys-1522 contains the radiolabel. This residue has been shown by site-directed mutagenesis to be essential for rat LAR activity (Pot, D. A., Woodford, T. A., Remboutsika, E., Haun, R. S., and Dixon, J. E. (1991) J. Biol. Chem. 266, 19688-19696). Iodoacetate reacts only with the first domain of this double domain PTPase. These results, for the first time, directly identify the highly reactive cysteine residue at the active site of a PTPase and highlight the ability of this residue to participate as a nucleophile in the hydrolysis of phosphate from tyrosine. |
| doi_str_mv | 10.1016/S0021-9258(18)48470-0 |
| format | Article |
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The data indicate that iodoacetate binds at the active site of the enzyme with a stoichiometry of 0.8 mol of iodoacetate bound per mol of rat LAR. A single [14C]iodoacetate-labeled peptide was isolated following endoproteinase Lys-C digestion of the radiolabeled PTPase. Sequence analysis of the active site labeled peptide demonstrates that Cys-1522 contains the radiolabel. This residue has been shown by site-directed mutagenesis to be essential for rat LAR activity (Pot, D. A., Woodford, T. A., Remboutsika, E., Haun, R. S., and Dixon, J. E. (1991) J. Biol. Chem. 266, 19688-19696). Iodoacetate reacts only with the first domain of this double domain PTPase. These results, for the first time, directly identify the highly reactive cysteine residue at the active site of a PTPase and highlight the ability of this residue to participate as a nucleophile in the hydrolysis of phosphate from tyrosine.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(18)48470-0</identifier><identifier>PMID: 1730581</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Analytical, structural and metabolic biochemistry ; Aniline Compounds - pharmacology ; Animals ; Binding Sites ; Binding, Competitive ; Biological and medical sciences ; Chromatography, High Pressure Liquid ; Enzyme Activation - drug effects ; Enzymes and enzyme inhibitors ; Fundamental and applied biological sciences. Psychology ; Glycoproteins - antagonists & inhibitors ; Glycoproteins - metabolism ; Hydrolases ; Iodoacetates - pharmacology ; Iodoacetic Acid ; LAR protein ; Molecular Sequence Data ; Organophosphorus Compounds - pharmacology ; Protein Tyrosine Phosphatases - antagonists & inhibitors ; Protein Tyrosine Phosphatases - genetics ; Protein Tyrosine Phosphatases - metabolism ; protein-tyrosine phosphatase ; Rats ; Receptor-Like Protein Tyrosine Phosphatases, Class 4 ; Receptors, Cell Surface ; Recombinant Proteins - antagonists & inhibitors ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism</subject><ispartof>The Journal of biological chemistry, 1992-01, Vol.267 (1), p.140-143</ispartof><rights>1992 © 1992 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c559t-ee2ef56fefb000578f22dd88efcb092b1f07c108aab6f3755f8bc80136364bbe3</citedby><cites>FETCH-LOGICAL-c559t-ee2ef56fefb000578f22dd88efcb092b1f07c108aab6f3755f8bc80136364bbe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5159655$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1730581$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pot, D A</creatorcontrib><creatorcontrib>Dixon, J E</creatorcontrib><title>Active site labeling of a receptor-like protein tyrosine phosphatase</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The inactivation of the cytoplasmic domain of rat LAR, a receptor-like protein tyrosine phosphatase (PTPase), by iodoacetate and not by iodoacetamide suggested that iodoacetate interacts in a highly selective manner with the enzyme. The data indicate that iodoacetate binds at the active site of the enzyme with a stoichiometry of 0.8 mol of iodoacetate bound per mol of rat LAR. A single [14C]iodoacetate-labeled peptide was isolated following endoproteinase Lys-C digestion of the radiolabeled PTPase. Sequence analysis of the active site labeled peptide demonstrates that Cys-1522 contains the radiolabel. This residue has been shown by site-directed mutagenesis to be essential for rat LAR activity (Pot, D. A., Woodford, T. A., Remboutsika, E., Haun, R. S., and Dixon, J. E. (1991) J. Biol. Chem. 266, 19688-19696). Iodoacetate reacts only with the first domain of this double domain PTPase. These results, for the first time, directly identify the highly reactive cysteine residue at the active site of a PTPase and highlight the ability of this residue to participate as a nucleophile in the hydrolysis of phosphate from tyrosine.</description><subject>Amino Acid Sequence</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Aniline Compounds - pharmacology</subject><subject>Animals</subject><subject>Binding Sites</subject><subject>Binding, Competitive</subject><subject>Biological and medical sciences</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Enzyme Activation - drug effects</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glycoproteins - antagonists & inhibitors</subject><subject>Glycoproteins - metabolism</subject><subject>Hydrolases</subject><subject>Iodoacetates - pharmacology</subject><subject>Iodoacetic Acid</subject><subject>LAR protein</subject><subject>Molecular Sequence Data</subject><subject>Organophosphorus Compounds - pharmacology</subject><subject>Protein Tyrosine Phosphatases - antagonists & inhibitors</subject><subject>Protein Tyrosine Phosphatases - genetics</subject><subject>Protein Tyrosine Phosphatases - metabolism</subject><subject>protein-tyrosine phosphatase</subject><subject>Rats</subject><subject>Receptor-Like Protein Tyrosine Phosphatases, Class 4</subject><subject>Receptors, Cell Surface</subject><subject>Recombinant Proteins - antagonists & inhibitors</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1LxDAQhoMoun78BKGCiB6qmbZp05OI3yB4UMFbSNKJjXbbNckq_nuzdtGjcwnMPJN5eQjZBXoMFMqTB0ozSOuM8UPgRwUvKprSFTIByvM0Z_C8Sia_yAbZ9P6VxipqWCfrUOWUcZiQizMd7Acm3gZMOqmws_1LMphEJg41zsLg0s6-YTJzQ0DbJ-HLDd72sdEOftbKID1ukzUjO487y3eLPF1dPp7fpHf317fnZ3epZqwOKWKGhpUGjYpBWMVNljUN52i0onWmwNBKx_hSqtLkFWOGK80p5GVeFkphvkUOxn9jmPc5-iCm1mvsOtnjMPcCSlpDXvEIshHUMax3aMTM2al0XwKoWNgTP_bEQo0ALn7sCRr3dpcH5mqKzd_WqCvO95dz6bXsjJO9tv4XY8DqkrGI7Y1Ya1_aT-tQKDvoFqciKysBAorFqdORwSjsw6ITXlvsNTaR10E0g_0n7DfAa5co</recordid><startdate>19920105</startdate><enddate>19920105</enddate><creator>Pot, D A</creator><creator>Dixon, J E</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>8FD</scope><scope>FR3</scope><scope>M7Z</scope><scope>P64</scope></search><sort><creationdate>19920105</creationdate><title>Active site labeling of a receptor-like protein tyrosine phosphatase</title><author>Pot, D A ; Dixon, J E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c559t-ee2ef56fefb000578f22dd88efcb092b1f07c108aab6f3755f8bc80136364bbe3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Amino Acid Sequence</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Aniline Compounds - pharmacology</topic><topic>Animals</topic><topic>Binding Sites</topic><topic>Binding, Competitive</topic><topic>Biological and medical sciences</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Enzyme Activation - drug effects</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glycoproteins - antagonists & inhibitors</topic><topic>Glycoproteins - metabolism</topic><topic>Hydrolases</topic><topic>Iodoacetates - pharmacology</topic><topic>Iodoacetic Acid</topic><topic>LAR protein</topic><topic>Molecular Sequence Data</topic><topic>Organophosphorus Compounds - pharmacology</topic><topic>Protein Tyrosine Phosphatases - antagonists & inhibitors</topic><topic>Protein Tyrosine Phosphatases - genetics</topic><topic>Protein Tyrosine Phosphatases - metabolism</topic><topic>protein-tyrosine phosphatase</topic><topic>Rats</topic><topic>Receptor-Like Protein Tyrosine Phosphatases, Class 4</topic><topic>Receptors, Cell Surface</topic><topic>Recombinant Proteins - antagonists & inhibitors</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pot, D A</creatorcontrib><creatorcontrib>Dixon, J E</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 1</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pot, D A</au><au>Dixon, J E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Active site labeling of a receptor-like protein tyrosine phosphatase</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1992-01-05</date><risdate>1992</risdate><volume>267</volume><issue>1</issue><spage>140</spage><epage>143</epage><pages>140-143</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>The inactivation of the cytoplasmic domain of rat LAR, a receptor-like protein tyrosine phosphatase (PTPase), by iodoacetate and not by iodoacetamide suggested that iodoacetate interacts in a highly selective manner with the enzyme. The data indicate that iodoacetate binds at the active site of the enzyme with a stoichiometry of 0.8 mol of iodoacetate bound per mol of rat LAR. A single [14C]iodoacetate-labeled peptide was isolated following endoproteinase Lys-C digestion of the radiolabeled PTPase. Sequence analysis of the active site labeled peptide demonstrates that Cys-1522 contains the radiolabel. This residue has been shown by site-directed mutagenesis to be essential for rat LAR activity (Pot, D. A., Woodford, T. A., Remboutsika, E., Haun, R. S., and Dixon, J. E. (1991) J. Biol. Chem. 266, 19688-19696). Iodoacetate reacts only with the first domain of this double domain PTPase. 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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Amino Acid Sequence Analytical, structural and metabolic biochemistry Aniline Compounds - pharmacology Animals Binding Sites Binding, Competitive Biological and medical sciences Chromatography, High Pressure Liquid Enzyme Activation - drug effects Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology Glycoproteins - antagonists & inhibitors Glycoproteins - metabolism Hydrolases Iodoacetates - pharmacology Iodoacetic Acid LAR protein Molecular Sequence Data Organophosphorus Compounds - pharmacology Protein Tyrosine Phosphatases - antagonists & inhibitors Protein Tyrosine Phosphatases - genetics Protein Tyrosine Phosphatases - metabolism protein-tyrosine phosphatase Rats Receptor-Like Protein Tyrosine Phosphatases, Class 4 Receptors, Cell Surface Recombinant Proteins - antagonists & inhibitors Recombinant Proteins - genetics Recombinant Proteins - metabolism |
| title | Active site labeling of a receptor-like protein tyrosine phosphatase |
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