Elevated carbohydrate phosphotransferase activity in human hepatoma and phosphorylation of cathepsin D.
To determine the cause of the increased content of carbohydrate-bound phosphate in tumour lysosomal hydrolases, the activity and kinetics in human hepatocellular carcinoma of two enzymes involved in the formation of mannose-6-phosphate in lysosomal hydrolases UDP-GlcNAc: lysosomal enzyme GlcNAc alpha l-phosphotransferase (GlcNAc-phosphotransferase) and phosphodiester glycosidase were studied. The activity level of the phosphotransferase with artificial and natural substrates was elevated (P less than 0.025 and P less than 0.001, respectively) in hepatoma compared to that in uninvolved tissue, while the phosphodiester glycosidase of hepatoma was at a level similar to that of the uninvolved tissue. To verify a previous observation that cathepsin D of human hepatoma contained increased GlcNAc-phosphomannose, the protease was examined for carbohydrate phosphorylation by the GlcNAc-phosphotransferase. The protease from normal human liver was much more phosphorylated than hepatoma protease, confirming the previous observation. The predominant phosphorylation of the protease occurred in one of two major heavy subunits, with some phosphorylation in one of two minor light subunits.
Ohhira
M
M
Biochemistry Laboratory, Hokkaido University School of Medicine, Sapporo, Japan.
Gasa
S
S
Makita
A
A
Sekiya
C
C
Namiki
M
M
D000230:Adenocarcinoma / Q000201:enzymology; D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D002402:Cathepsin D / Q000302:isolation & purification / Q000378:metabolism*; D005706:Gallbladder Neoplasms / Q000201:enzymology; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008103:Liver Cirrhosis / Q000201:enzymology; D008113:Liver Neoplasms / Q000201:enzymology*; D008970:Molecular Weight; D010750:Phosphoproteins / Q000302:isolation & purification; D010766:Phosphorylation; D010770:Phosphotransferases / Q000302:isolation & purification / Q000378:metabolism*; D017855:Transferases (Other Substituted Phosphate Groups)
D010750:Phosphoproteins; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); C030012:UDP-N-acetylglucosamine-lysosomal-enzyme-N-acetylglucosaminephosphotransferase; D002402:Cathepsin D