Purification and characterization of N-acetyl-beta-D-hexosaminidase in different anatomical regions of the adult rat epididymis.
The purpose of the present study was to purify and kinetically characterize N-acetyl-beta-D-hexosaminidases A and B (EC 3.2.1.52) from the caput, corpus and caudal regions of the adult rat epididymis. The molecular mass of the purified native enzyme was approximately 250,000 and approximately 223,000 daltons for the A and B isozymes, with a subunit molecular mass of approximately 63,000 and approximately 56,000 daltons, as determined by size exclusion chromatography and gel electrophoresis under reducing conditions. The apparent Michaelis-Menten constant and maximum velocity values were 0.60, 1.55 and 0.68 mM and 0.54, 3.20 and 2.30 microM/min./mg protein for the enzyme purified from the caput, corpus and caudal regions, respectively. These values were determined by using p-nitrophenyl-N-acetyl-beta-D-glucosaminide as the substrate. These data suggest that the enzyme may be more active in the corpus region of the epididymis than in the caput and caudal regions.
Hall
J C
JC
Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Kochins
J G
JG
Perez
F M
FM
D000818:Animals; D002850:Chromatography, Gel; D004795:Enzyme Stability; D004822:Epididymis / Q000201:enzymology*; D006358:Hot Temperature; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution; D001619:beta-N-Acetylhexosaminidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D007527:Isoenzymes; D001619:beta-N-Acetylhexosaminidases