TY - JOUR
T1 - Electrophoretic separation of neutral and acid β-glucosidase isozymes in human tissues
AU - Shafit-Zagardo, B.
AU - Devine, E. A.
AU - Desnick, R. J.
N1 - Funding Information:
We wish to thank Dr. R. Schaler for tissue specimens and Drs. D.F. Bishop and G.A. Grabowski for helpful experimental advice. This work was supported by a grant (1-438) from the National Foundation-March of Dimes. B.S.-Z. and E.A.D. are recipients of NIH pre-and post-doctoral fellowships, respectively (HD 07105), and R.J.D. is the recipient of an NIH Research Career Development Award (K01 AM 00451).
PY - 1980
Y1 - 1980
N2 - An electrophoretic system using cellulose acetate has been developed for the resolution of β-glucosidase isozymes (β-D-glucoside glucohydrolase, EC 3.2.1.21 and D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45) in human tissue homogenates. Electrophoresis of homogenates from normal and Type I Gaucher disease tissues revealed two fluorescent bands of β-glucosidase activity which corresponded to the acid and neutral isozymes separated by concanavalin A-Sepharose chromatography. The acid isozyme had only β-glucosidase activity, whereas the neutral isozyme also exhibited α-L-arabinosidase (α-L-arabino-furanoside arabinofuranohydrolase, EC 3.2.1.55), β-D-galactosidase (β-D galactoside galactohydrolase, , EC 3.2.1.23) and β-D-xylosidase (1,4-β-D-xylan xylohydrolase, EC 3.2.1.37) activities, using the appropriate 4-methylumbelliferyl glycoside. In homogenates of cultured skin fibroblasts, only the acid isozyme was observed which co-electrophoresed with the acidic activity in other tissue homogenates. The acidic activity in tissue and fibroblast homogenates from Type I Gaucher disease appeared to co-electrophorese with the acid isozyme in normal tissues, but had markedly reduced activity.
AB - An electrophoretic system using cellulose acetate has been developed for the resolution of β-glucosidase isozymes (β-D-glucoside glucohydrolase, EC 3.2.1.21 and D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45) in human tissue homogenates. Electrophoresis of homogenates from normal and Type I Gaucher disease tissues revealed two fluorescent bands of β-glucosidase activity which corresponded to the acid and neutral isozymes separated by concanavalin A-Sepharose chromatography. The acid isozyme had only β-glucosidase activity, whereas the neutral isozyme also exhibited α-L-arabinosidase (α-L-arabino-furanoside arabinofuranohydrolase, EC 3.2.1.55), β-D-galactosidase (β-D galactoside galactohydrolase, , EC 3.2.1.23) and β-D-xylosidase (1,4-β-D-xylan xylohydrolase, EC 3.2.1.37) activities, using the appropriate 4-methylumbelliferyl glycoside. In homogenates of cultured skin fibroblasts, only the acid isozyme was observed which co-electrophoresed with the acidic activity in other tissue homogenates. The acidic activity in tissue and fibroblast homogenates from Type I Gaucher disease appeared to co-electrophorese with the acid isozyme in normal tissues, but had markedly reduced activity.
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U2 - 10.1016/0005-2744(80)90235-1
DO - 10.1016/0005-2744(80)90235-1
M3 - Article
C2 - 6773581
AN - SCOPUS:0019120325
SN - 0006-3002
VL - 614
SP - 459
EP - 465
JO - Biochimica et biophysica acta
JF - Biochimica et biophysica acta
IS - 2
ER -
