HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Orru, S. Articles by Esposito, C. Search for Related Content PubMed PubMed Citation Articles by Orru, S. Articles by Esposito, C. Social Bookmarking                   What's this?

Identification of tissue transglutaminase-reactive lysine residues in glyceraldehyde-3-phosphate dehydrogenase

¹ Dipartimento di Chimica, Università di Salerno, Salerno, Italy
² Centro di Studio di Biocristallografia, CNR, Naples, Italy
³ Dipartimento di Chimica Organica e Biochimica, Università di Napoli Federico II, Naples, Italy

Reprint requests to: Prof. Carla Esposito, Dipartimento di Chimica, Università di Salerno, Via S. Allende, 84081 Baronissi, Salerno, Italy; e-mail: esposito{at}mbox.chem.unisa.it; fax:39-089-965296.

Polyglutamine domains are excellent substrates for tissue transglutaminase resulting in the formation of cross-links with polypeptides containing lysyl residues. This finding suggests that tissue transglutaminase may play a role in the pathology of neurodegenerative diseases associated with polyglutamine expansion. The glycolytic enzyme GAPDH previously was shown to tightly bind several proteins involved in such diseases. The present study confirms that GAPDH is an in vitro lysyl donor substrate of tissue transglutaminase. A dansylated glutamine-containing peptide was used as probe for labeling the amino-donor sites. SDS gel electrophoresis of a time-course reaction mixture revealed the presence of both fluorescent GAPDH monomers and high molecular weight polymers. Western blot analysis performed using antitransglutaminase antibodies reveals that tissue transglutaminase takes part in the formation of heteropolymers. The reactive amino-donor sites were identified using mass spectrometry. Here, we report that of the 26 lysines present in GAPDH, K191, K268, and K331 were the only amino-donor residues modified by tissue transglutaminase.

Keywords: GAPDH; lysine residues; mass spectrometry; tissue transglutaminase

Abbreviations: Asp-N, endoproteinase Asp-N • DNS, dansyl group, ESI-MS, electrospray mass spectrometry • HD, Huntington disease • LC/MS, liquid chromatography/mass spectrometry • MALDI-MS, matrix-assisted laser desorption ionization mass spectrometry • Qn, polyglutamine repeat containing n Q residues • RP-HPLC, reverse phase high pressure liquid chromatography • SubP, substance P • TG, transglutaminase • TIC, total ion current • tTG, guinea pig liver tissue transglutaminase

CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. L. Griffin, C. K. Cemal, and M. A. Pook Defining a metabolic phenotype in the brain of a transgenic mouse model of spinocerebellar ataxia 3 Physiol Genomics, February 13, 2004; 16(3): 334 - 340. [Abstract] [Full Text] [PDF]

				S. Orru, I. Caputo, A. D'Amato, M. Ruoppolo, and C. Esposito Proteomics Identification of Acyl-acceptor and Acyl-donor Substrates for Transglutaminase in a Human Intestinal Epithelial Cell Line: IMPLICATIONS FOR CELIAC DISEASE J. Biol. Chem., August 22, 2003; 278(34): 31766 - 31773. [Abstract] [Full Text] [PDF]

				M. Ruoppolo, S. Orru, S. Francese, I. Caputo, and C. Esposito Structural characterization of transglutaminase-catalyzed cross-linking between glyceraldehyde 3-phosphate dehydrogenase and polyglutamine repeats Protein Sci., January 1, 2003; 12(1): 170 - 179. [Abstract] [Full Text] [PDF]