Identification of tissue transglutaminase-reactive lysine residues in glyceraldehyde-3-phosphate dehydrogenase(9804 views) Orru S, Ruoppolo M, Francese S, Vitagliano L, Marino G, Esposito C
Protein Sci (ISSN: 0961-8368, 1469-896xelectronic), 2002 Jan; 11(1): 137-146.
Keywords: Gapdh, Lysine Residues, Mass Spectrometry, Tissue Transglutaminase, Enzyme Antibody, Glyceraldehyde 3 Phosphate Dehydrogenase, Polyglutamine, Protein Glutamine Gamma Glutamyltransferase, Article, Degenerative Disease, Enzyme Analysis, Enzyme Modification, Enzyme Substrate, Fluorescence Analysis, Molecular Probe, Polyacrylamide Gel Electrophoresis, Priority Journal, Protein Cross Linking, Western Blotting, Animals, Glyceraldehyde-3-Phosphate Dehydrogenases, Microscopy, Models, Peptides, Protein Binding, Protein Conformation, Protein Structure, Secondary, Tertiary, Rabbits, Matrix-Assisted Laser Desorption-Ionization, Substrate Specificity, Swine, Time Factors,
Affiliations: *** IBB - CNR ***
Dipartimento di Chimica, Università di Salerno, Salerno, Italy
Centro di Studio di Biocristallografia, CNR, Naples, Italy
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Paulson, H. L., Perez, M. K., Trottier, Y., Trojanowski, J. Q., Subramony, S. H., Das, S. S., Vig, P., Pittman, R. N., Intranuclear inclusions of expanded polyglutamine protein in spinocerebellar ataxia type 3 (1997) Neuron, 19, pp. 333-344
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Perutz, M. F., Johnson, T., Suzuki, M., Finch, J. T., Glutamine repeats as polar zippers: Their possible role in inherited neurodegenerative diseases (1994) Proc. Natl. Acad. Sci., 91, pp. 5355-5358
Saunders, P. A., Chalecka-Franaszek, E., Chuang, D. M., Subcellular distribution of glyceraldehyde 3-phosphate dehydrogenase in cerebellar granule cells undergoing cytosine arabinoside-induced apoptosis (1997) J. Neurochem., 69, pp. 1820-1828
Sawa, A., Khan, A. A., Hester, L. D., Snyder, S. H., Glyceraldehyde 3-phosphate dehydrogenase: Nuclear translocation in neuronal and nonneuronal cell death (1997) Proc. Natl. Acad. Sci., 90, pp. 549-558
Sirover, M. A., New insights into an old protein: The functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase (1999) Biochem. Biophys. Acta, 1432, pp. 159-184
Steinert, P. M., Candi, E., Tarcsa, E., Marekov, L. N., Sette, M., Paci, M., Ciani, B., Melino, G., Transglutaminase crosslinking and structural studies of the human small proline rich protein (1999) Cell Death Differ., 6, pp. 916-930
Walker, J. E., Carne, A. F., Runswick, M. J., Bridgen, J., Harris, J. I., D-glyceraldehyde 3-phosphate dehydrogenase. Complete amino acid sequence of the enzyme from Bacillus stearothermophilus (1980) Eur. J. Biochem., 108, pp. 549-565
Identification of tissue transglutaminase-reactive lysine residues in glyceraldehyde-3-phosphate dehydrogenase
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. SIDS 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 trans glutaminase.
Identification of tissue transglutaminase-reactive lysine residues in glyceraldehyde-3-phosphate dehydrogenase
No results.
Identification of tissue transglutaminase-reactive lysine residues in glyceraldehyde-3-phosphate dehydrogenase