The separation of bovine brain beta-N-acetyl-D-hexosaminidases. Abnormal gel-filtration behaviour of beta-N-acetyl-D-glucosaminidase C

• Published in: Biochemical journal 1975-11, Vol.151 (2), p.257-261
• Main Authors: Overdijk, B, van der Kroef, W M, Veltkamp, W A, Hooghwinkel, G J
• Format: Article
• Language: English
• Subjects: Acetylgalactosamine - metabolism; Acetylglucosaminidase - isolation & purification; Animals; Brain - enzymology; Cattle; Chromatography, Gel; Chromatography, Ion Exchange; Electrophoresis; Hexosaminidases - isolation & purification; Hexosaminidases - metabolism
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/bj1510257

Bovine brain tissue was extracted and the 50 000g supernatant was separated by electrophoresis, DEAE-Sephadex chromatography and gel filtration on Sephadex G-200 and Bio-Gel P-200. The electrophoretic separation showed that the beta-N-acetyl-D-hexosaminidases (hexosaminidases) of bovine brain tissue were composed of four different fractions. Two fractions (A and B) exerted both glucosaminidase and galactosaminidase activity, a third fraction (C) showed only glucosaminidase activity, whereas a fourth form (D) with specificity towards the galactosaminide moiety was found to be present. DEAE-Sephadex chromatography at pH 7.0 showed that the B form was eluted with the void volume, whereas the A and D forms could be eluted in one peak by raising that salt concentration. The C form could not be detected in the eluate. Gel filtration on Sephadex G-200 showed that the B, A and D forms had almost equal molecular weights. In this case also the C form could not be detected in the column eluates. Gel filtration on Bio-Gel P-200 revealed that the C form was eluted with the void volume.