Abstract
Human serum α-l-fucosidase has been purified 241 200-fold with 35% yield by an affinity Chromatographie procedure utilizing agarose-ε{lunate}-aminocaproyl-fucosamine. Isoelectric focusing of the purified enzyme indicated the presence of several forms, with the form at pI 5.0 comprising the majority of the activity. Assay of the purified α-l-fucosidase showed only trace amounts of contaminating glycosidases present, with β-galactosidase being the largest contamnant (0.5% by activity). Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated the presence of two subunits with very similar molecular weights (56500 and 54000). Using the p-nitrophenyl substrate, the purified serum α-l-fucosidase had an apparent Michaelis constant of 0.52 mM and a broad pH optimum centered around pH 4.8 with a second, minor optimum at pH 6.1. Gel filtration on Sepharose 6-B indicated an apparent molecular weight of 296 000 ± 30 000. Preincubation with antibodies made previously against purified liver α-l-fucosidase led to quantitative immuno precipitation of the purified serum α-l-fucosidase. Assay of the purified serum α-l-fucosidase for sialic acid indicated the presence of 1.7 μg sialic acid per 100 μg enzyme, about twice that previously found for the purified liver enzyme.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 133-140 |
| Number of pages | 8 |
| Journal | Clinica Chimica Acta |
| Volume | 82 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Jan 2 1978 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
- Clinical Biochemistry
Cite this
Human serum α-l-fucosidase. / Alhadeff, Jack A.; Janowsky, Aaron.
In: Clinica Chimica Acta, Vol. 82, No. 1-2, 02.01.1978, p. 133-140.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Human serum α-l-fucosidase
AU - Alhadeff, Jack A.
AU - Janowsky, Aaron
PY - 1978/1/2
Y1 - 1978/1/2
N2 - Human serum α-l-fucosidase has been purified 241 200-fold with 35% yield by an affinity Chromatographie procedure utilizing agarose-ε{lunate}-aminocaproyl-fucosamine. Isoelectric focusing of the purified enzyme indicated the presence of several forms, with the form at pI 5.0 comprising the majority of the activity. Assay of the purified α-l-fucosidase showed only trace amounts of contaminating glycosidases present, with β-galactosidase being the largest contamnant (0.5% by activity). Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated the presence of two subunits with very similar molecular weights (56500 and 54000). Using the p-nitrophenyl substrate, the purified serum α-l-fucosidase had an apparent Michaelis constant of 0.52 mM and a broad pH optimum centered around pH 4.8 with a second, minor optimum at pH 6.1. Gel filtration on Sepharose 6-B indicated an apparent molecular weight of 296 000 ± 30 000. Preincubation with antibodies made previously against purified liver α-l-fucosidase led to quantitative immuno precipitation of the purified serum α-l-fucosidase. Assay of the purified serum α-l-fucosidase for sialic acid indicated the presence of 1.7 μg sialic acid per 100 μg enzyme, about twice that previously found for the purified liver enzyme.
AB - Human serum α-l-fucosidase has been purified 241 200-fold with 35% yield by an affinity Chromatographie procedure utilizing agarose-ε{lunate}-aminocaproyl-fucosamine. Isoelectric focusing of the purified enzyme indicated the presence of several forms, with the form at pI 5.0 comprising the majority of the activity. Assay of the purified α-l-fucosidase showed only trace amounts of contaminating glycosidases present, with β-galactosidase being the largest contamnant (0.5% by activity). Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated the presence of two subunits with very similar molecular weights (56500 and 54000). Using the p-nitrophenyl substrate, the purified serum α-l-fucosidase had an apparent Michaelis constant of 0.52 mM and a broad pH optimum centered around pH 4.8 with a second, minor optimum at pH 6.1. Gel filtration on Sepharose 6-B indicated an apparent molecular weight of 296 000 ± 30 000. Preincubation with antibodies made previously against purified liver α-l-fucosidase led to quantitative immuno precipitation of the purified serum α-l-fucosidase. Assay of the purified serum α-l-fucosidase for sialic acid indicated the presence of 1.7 μg sialic acid per 100 μg enzyme, about twice that previously found for the purified liver enzyme.
UR - http://www.scopus.com/inward/record.url?scp=0017918327&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0017918327&partnerID=8YFLogxK
U2 - 10.1016/0009-8981(78)90036-0
DO - 10.1016/0009-8981(78)90036-0
M3 - Article
C2 - 618676
AN - SCOPUS:0017918327
VL - 82
SP - 133
EP - 140
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
SN - 0009-8981
IS - 1-2
ER -