Advertisement

FAB Mass Spectrometric Detection of ε( γ -Glutamyl)Lysine Crosslinks and ( γ -Glutamyl)Polyamine Derivatives Produced by Transglutaminase in Vitro

  • P. Pucci
  • G. Marino
  • A. Malorni
  • S. Metafora
  • C. Esposito
  • R. Porta
Part of the Advances in Experimental Medicine and Biology book series (NATO ASI F, volume 231)

Abstract

The identification of both acyl donor and acceptor substrates, as well as the detection of ε( γ -glutamyl)lysine crosslinks and ( γ-glutamyl) amine derivatives, are some of the most important aspects in the field of the transglutaminase (TGase)-mediated reactions1–5. These reactions have been revealed so far using both direct and indirect methodologies. The main direct method is based upon chromatographic isolation and identification of the isopeptide ε( γ -glutamyl)lysine or of ( γ -glutamyl)amine derivatives after exhaustive proteolytic digestion of the reaction products. On the other hand, the detection of either polymers or radioactive amine-protein adducts by SDS-PAGE, together with the protein crosslinking inhibition by small molecular weight amines, represent the most commonly used indirect methodologies. However, both procedures suffer from severe limitations since a single analytical system does not yield unambiguous results.

Keywords

Lysine Residue Fast Atom Bombardment Acyl Donor Acceptor Substrate Glutamine Residue 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    H. G. Williams-Ashman and Z. N. Canellakis, Transglutaminase-mediated covalent attachment of polyamines to proteins: mechanisms and potential physiological significance, Physiol. Chem. Phys. 12:457 (1980).PubMedGoogle Scholar
  2. 2.
    J. E. Folk, Mechanisms and basis for specificity of transglutaminase-catalyzed epsilon(gamma-glutamyl)lysine bond formation, in: “Advances in Enzymology and Related Areas in Molecular Biology,” Meister, A. ed., Wiley, New York (1983).Google Scholar
  3. 3.
    L. Lorand and S. M. Conrad, Transglutaminases, Mol. Cell. Biochem. 58:9 (1984).PubMedCrossRefGoogle Scholar
  4. 4.
    M. Griffin and J. Wilson, Detection of ε(γ-glutamyl)lysine, Mol. Cell. Biochem. 58:37 (1984).PubMedCrossRefGoogle Scholar
  5. 5.
    A. G. Loewy, The Nε-(γ-glutamic)lysine cross-link: method of analysis, occurrence in extracellular and cellular proteins, in: “Meth. Enzymol.” 107:241, Academic Press, Inc, New York (1984).Google Scholar
  6. 6.
    A. Dell and M. Panico, Fast Atom Bombardment mass spectrometry of biomolecules, in: “Mass Spectrometry in Biomedical Research,” S. Gaskell, ed., J. Wiley and sons, Chichester (1986).Google Scholar
  7. 7.
    S. Nakanishi, A. Inoue, T. Kita, M. Nakamura, A. C. Y. Chang, S. N. Cohen and S. Numa, Nucleotide sequence of cloned cDNA for bovine corticotropin-β-lipotropin precursor, Nature 278:423 (1979).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1988

Authors and Affiliations

  • P. Pucci
    • 1
  • G. Marino
    • 1
  • A. Malorni
    • 3
  • S. Metafora
    • 4
  • C. Esposito
    • 2
  • R. Porta
    • 2
  1. 1.Department of Organic and Biological Chemistry, 1st Medical SchoolUniversity of NaplesNaplesItaly
  2. 2.Institute of Chemistry and Biological Chemistry, 1st Medical SchoolUniversity of NaplesNaplesItaly
  3. 3.Institutes of Chemistry of Molecules of Biological InterestCNRArco Felice, NaplesItaly
  4. 4.Institutes of Protein Biochemistry and EnzymologyCNRNaplesItaly

Personalised recommendations