Bovine testicular beta-galactosidase: purification of enzyme fractions that exhibit high affinity for phosphomannosyl receptors.
An improved method is described for the preparation of bovine testicular beta-galactosidase that allows the isolation of enzyme fractions that bind avidly to phosphomannosyl receptors. The procedure permits removal of a contaminating beta-hexosaminidase and yields nearly homogeneous beta-galactosidase. Enzyme eluted from DEAE-Sephacel was arbitrarily divided into pools that exhibited differing ability to bind phosphomannosyl receptors. A high binding fraction was rapidly assimilated by cultured cells and bound to both low and high molecular weight phosphomannosyl receptors. Carbohydrate analysis of the high binding fraction indicates an average content of one complex and one high mannose oligosaccharide chain per molecule and an average mannose 6-phosphate content of two residues per molecule. However, electrofocusing studies indicated that all the fractions were heterogeneous with respect to sialic acid and phosphate content. The purification procedure also provides highly purified beta-galactosidase suitable for removing beta-galactosidase residues from a variety of complex carbohydrates.
Distler
J J
JJ
Rackham Arthritis Research Unit, University of Michigan Medical School, Ann Arbor 48109.
Guo
J F
JF
Sahagian
G G
GG
Jourdian
G W
GW
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000379:methods; D003636:DEAE-Cellulose; D005696:Galactosidases / Q000302:isolation & purification*; D006596:Hexosaminidases / Q000032:analysis; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000032:analysis; D008297:Male; D008970:Molecular Weight; D009439:Neuraminidase / Q000032:analysis; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear; D012685:Sepharose / Q000031:analogs & derivatives; D013737:Testis / Q000201:enzymology*; D001616:beta-Galactosidase / Q000302:isolation & purification* / Q000378:metabolism; D048828:beta-N-Acetyl-Galactosaminidase
D002241:Carbohydrates; D007527:Isoenzymes; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; C034304:cation-dependent mannose-6-phosphate receptor; C028326:concanavalin A-sepharose; D012685:Sepharose; D003636:DEAE-Cellulose; D005696:Galactosidases; D006596:Hexosaminidases; D009439:Neuraminidase; D001616:beta-Galactosidase; D048828:beta-N-Acetyl-Galactosaminidase