Comparison of the peptide and saccharide moieties of gamma-glutamyltransferase isolated from neoplastic and non-neoplastic human liver tissue.
gamma-Glutamyltransferase from human hepatoma and the surrounding non-neoplastic liver tissue was purified by immunoaffinity column chromatography and characterized with regard to molecular weight, isoelectric point (pI), amino acid composition, hexosamine content and affinity for various lectins. Both enzymes showed the same molecular weight (the heavy subunit 64 000; the light subunit 26 000) and pIs (3.7-3.9) on SDS-polyacrylamide gel electrophoresis and isoelectric focusing in polyacrylamide gel, respectively. After neuraminidase treatment, the pIs for both enzymes shifted to a more alkaline pH (pI 5.7). Both enzyme preparations exhibited similar amino acid compositions; however, the glucosamine content of the hepatoma enzyme was 362 nmol/mg protein, about 3-fold higher than that of the enzyme isolated protein from the non-neoplastic tissue. Binding of the two enzymes to lectins revealed that less of the hepatoma enzyme bound to Sepharose-conjugated wheat germ agglutinin, erythroagglutinating phytohemagglutinin and Ricinus communis agglutinin. These results suggest that the two enzymes possess similar peptide moieties and degree of sialylation, but differ with respect to other aspects of their heterosaccharide moieties.
Tsuchida
S
S
Yamazaki
T
T
Camba
E M
EM
Morita
T
T
Matsue
H
H
Yoshida
Y
Y
Sato
K
K
D000117:Acetylglucosamine / Q000494:pharmacology; D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis*; D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D003208:Concanavalin A / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D007525:Isoelectric Focusing; D037102:Lectins / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008113:Liver Neoplasms / Q000201:enzymology*; D008766:Methylmannosides / Q000494:pharmacology; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis*; D010835:Phytohemagglutinins / Q000378:metabolism; D037121:Plant Lectins; D014909:Wheat Germ Agglutinins; D005723:gamma-Glutamyltransferase / Q000032:analysis*
D000596:Amino Acids; D002241:Carbohydrates; D006595:Hexosamines; D037102:Lectins; D008766:Methylmannosides; D010455:Peptides; D010835:Phytohemagglutinins; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; C008466:methylmannoside; D005723:gamma-Glutamyltransferase; D000117:Acetylglucosamine