Characterisation of the cellular substrates for transglutaminase in normal liver and hepatocellular carcinoma

• Published in: Biochimica et biophysica acta Vol. 1033; no. 1; pp. 57 - 64
• Main Authors: Hand, David, Elliott, Barry M., Griffin, Martin
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 29.01.1990; Elsevier; North-Holland
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Calcium Chloride - pharmacology; Cytosol - enzymology; Dipeptides - metabolism; Egtazic Acid - pharmacology; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Liver - enzymology; Liver Neoplasms, Experimental - enzymology; Male; Methylamine; Methylamines - metabolism; Molecular Weight; Polyamine; Polyamines - metabolism; Rats; Transferases; Transglutaminase; Transglutaminases - metabolism; Tumor growth; ϵ-(γ-Glutamyl)lysine; Polyamine; Methylamine; Transglutaminase; ϵ-(γ-Glutamyl)lysine; Tumor growth; Carcinoma; Radiolabelling; Rat; Gel electrophoresis; Enzyme; Liver; Reaction product; Rodentia; HPLC chromatography; Normal; Characterization; Substrate; Absorption spectrometry; Vertebrata; Mammalia; Enzymatic activity; Tumor; Protein-glutamine γ-glutamyltransferase; Comparative study
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/0304-4165(90)90194-2

The transglutaminase-mediated incorporation of [ 14C]methylamine into tissue slices obtained from normal rat liver and diethylnitrosamine-induced hepatocellular carcinomas was used as a means of characterising the endogenous substrates of the transglutaminase enzymes present in these tissues. The amount of radiolabel incorporated was found to be similar in both tissues with the major radiolabelled protein identified as a high molecular weight polymer unable to traverse a 3.0% (w/v) acrylamide gel and with a molecular wieght of at least 5·10 6 Da. Measurement of the crosslink, ϵ-(γ-glutamyl)lysine, in the hepatocellular carcinoma and in normal liver indicated a 3-fold reduction in the levels found in tumour tissue when compared to normal liver. In contrast, the levels of covalently bound polyamines present in the hepatocellular carcinoma were found to be comparable or greater than those found in normal liver. Considering that there is a selective reduction (approx. 5-fold) in the activity of the cytosolic transglutaminase present in hepatocellular carcinomas with no change in the activity of the particulate enzyme (Hand et al. (1988) Biochim. Biophys. Acta 970, 137–145) these results suggest that the two enzymes may be differentially activated and that they may act on different substrates within the cell.