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Transglutaminase activity and N ϵ (γ‐glutamyl) lysine isopeptide levels during cell growth: An enzymic and immunological study
Author(s) -
Alaoui Said El,
Legastelois Stéphane,
Roch AnneMarie,
Chantepie Jacqueline,
Quash Gérard
Publication year - 1991
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910480212
Subject(s) - tissue transglutaminase , lysine , heterologous , biochemistry , monoclonal antibody , chemistry , peptide , microbiology and biotechnology , enzyme , antibody , biology , amino acid , immunology , gene
A monoclonal antibody (MAb) 81D1c2, which recognizes the Nϵ(γ‐glutamyl) lysine isopeptide produced by the action of transglutaminase activity was prepared. Its reactivity towards the homologous isopeptide was about 3‐fold greater than that with either Nα (α glutamyl) lysine (a naturally occuring heterologous dipeptide) or Nϵ(γ glutamyl) lysine, another heterologous peptide not described so far in naturally occuring proteins. When used in an immunohistochemical study on cells in culture derived from human carcinoma of the larynx (HEp2) and from chicken embryo cells (CEC), both fixed in acetone, this MAb detected Nϵ(γ glutamyl) lysine residues In the nucleus. The amount of Nϵ(γ glutamyl) lysine isopeptides follows closely transglutaminase activity during the lag phase of growth of both CEC and HEp2 cells. However, during exponential growth, the 2 parameters decrease concomitantly in HEp2 cells, whereas in CEC, transglutaminase activity increases but isopeptide bond levels drop. Compared with other reported methods for measuring isopeptides, this immunohistological approach permits the localization and at least the semi‐quantitative determination of Nϵ(γ glutamyl) lysine in cells in situ .