Purification and characterization of hexosaminidase from human uterine cervical carcinoma.
Normal human uterine cervical tissue and uterine cervical carcinoma tissue were collected and subjected to fractionation of hexosaminidase isoenzymes Hex A, Hex B, and Hex I using DEAE-cellulose anion-exchange chromatography. Hex A was found to be the major isoenzyme in control tissues, whereas Hex B was the major isoenzyme in carcinoma tissues. These two major isoenzyme fractions were first purified using heparin-Sepharose 4B affinity chromatography and then subjected to further purification on epsilon-ACMA-Sepharose 4B. The purified isoenzymes were found to be homogeneous by polyacrylamide gel electrophoresis. The Hex A and Hex B isoenzymes obtained from control and carcinoma patients were characterized. Hex A from control and carcinoma patients exhibited more or less similar properties. However, the Hex B isoenzyme from carcinoma patients exhibited some characteristic variations in pH, temperature, substrate concentration optima, and isoelectric point compared with the control. From these observations it may be inferred that altered properties of the Hex B isoenzyme fraction from carcinoma patients may be the reason for the elevated activity of hexosaminidase in carcinoma of the uterine cervix.
Bhuvarahamurthy
V
V
Department of Biochemistry, University of Madras, India.
Govindasamy
S
S
D005591:Chemical Fractionation; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D005260:Female; D054818:Hexosaminidase A; D054819:Hexosaminidase B; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008970:Molecular Weight; D012016:Reference Values; D013696:Temperature; D002583:Uterine Cervical Neoplasms / Q000737:chemistry* / Q000201:enzymology; D001619:beta-N-Acetylhexosaminidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*
D054818:Hexosaminidase A; D054819:Hexosaminidase B; D001619:beta-N-Acetylhexosaminidases