Structural characterization of transglutaminase-catalyzed cross-linking between glyceraldehyde 3-phosphate dehydrogenase and polyglutamine repeats

Margherita Ruoppolo, Stefania Orrù, Simona Francese, Ivana Caputo, Carla Esposito

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The accumulation of abnormal polyglutamine-containing protein aggregates within the cytosol and nuclei of affected neurons is a hallmark of the progressive neurodegenerative disorders caused by an elongated (CAG)n repeat in the genome. The polyglutamine domains are excellent substrates for the enzyme transglutaminase type 2 (tissue), resulting in the formation of cross-links with polypeptides containing lysyl groups. Enzymatic activity toward the Qn domains increases greatly upon lengthening of such Qn stretches (n > 40). Among the possible amine donors, the glycolytic enzyme glyceraldehyde-3-phosphate-dehydrogenase was shown to tightly bind several proteins involved in polyglutamine expansion diseases. Recently, the authors have shown that K191, K268, and K331, out of the 26 lysines present in glyceraldehyde-3-phosphate-dehydrogenase, are the reactive amine-donor sites forming cross-links with substance P, which bears the simplest Qn domain (n = 2). The present study reports that synthetic peptides of both pathological and nonpathological length (n = 43 and 17, respectively) form cross-links with the same K residues located in the C-terminal region of glyceraldehyde-3-phosphate-dehydrogenase. In addition, it is shown that extra K residues present in the C termini of glyceraldehyde-3-phosphate-dehydrogenase are susceptible to cross-linking in the presence of transglutaminase. The present results indicate a possible modulating effect of Qn stretches on tissue transglutaminase substrate specificity and mechanism of recognition.

Original languageEnglish
Pages (from-to)170-179
Number of pages10
JournalProtein Science
Volume12
Issue number1
DOIs
Publication statusPublished - Jan 1 2003

Fingerprint

Glyceraldehyde-3-Phosphate Dehydrogenases
Transglutaminases
Amines
Peptides
Substrates
Enzymes
Substance P
Substrate Specificity
Neurodegenerative Diseases
Cytosol
Lysine
Neurons
Genes
Genome
Tissue
polyglutamine
Proteins
transglutaminase 2

Keywords

  • Glyceraldehyde 3-phosphate dehydrogenase
  • Lysine residues
  • Mass spectrometry
  • Q disease
  • Transglutaminase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structural characterization of transglutaminase-catalyzed cross-linking between glyceraldehyde 3-phosphate dehydrogenase and polyglutamine repeats. / Ruoppolo, Margherita; Orrù, Stefania; Francese, Simona; Caputo, Ivana; Esposito, Carla.

In: Protein Science, Vol. 12, No. 1, 01.01.2003, p. 170-179.

Research output: Contribution to journalArticle

Ruoppolo, Margherita ; Orrù, Stefania ; Francese, Simona ; Caputo, Ivana ; Esposito, Carla. / Structural characterization of transglutaminase-catalyzed cross-linking between glyceraldehyde 3-phosphate dehydrogenase and polyglutamine repeats. In: Protein Science. 2003 ; Vol. 12, No. 1. pp. 170-179.
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