Bovine kidney beta-mannosidase: purification and characterization.
Lysosomal beta-mannosidase was purified 160,000-fold in 24% yield from bovine kidney by a four-step purification procedure, which included concanavalin A-Sepharose, immunoaffinity, TSK-butyl and h.p.l.c. cation-exchange chromatography. When analysed by SDS/PAGE and detected by Coomassie Blue or silver staining, the purified enzyme preparation consists of two prominent peptides (100 and 110 kDa) and a third minor peptide (84 kDa). These three peptides are immunologically related and are consistently associated with beta-mannosidase activity in all chromatographic steps. Removal of N-linked carbohydrate from the 84, 100 and 110 kDa peptides decreases their molecular sizes to 75, 86 and 91 kDa respectively. Bovine kidneys lacking beta-mannosidase, activity, acquired from calves affected with beta-mannosidosis, do not contain detectable quantities of the three beta-mannosidase peptides, as judged by monoclonal- and polyclonal-antibody reactivity.
Sopher
B L
BL
Department of Pathology, Michigan State University, East Lansing 48824.
Traviss
C E
CE
Cavanagh
K T
KT
Jones
M Z
MZ
Friderici
K H
KH
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D002418:Cattle Diseases; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008247:Lysosomes / Q000201:enzymology; D008361:Mannosidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D012016:Reference Values; D008363:alpha-Mannosidosis / Q000201:enzymology / Q000235:genetics / Q000662:veterinary; D044902:beta-Mannosidase
D002241:Carbohydrates; D008361:Mannosidases; D044902:beta-Mannosidase