S-Space College of Medicine/School of Medicine (의과대학/대학원) Dept. of Medicine (의학과) The Seoul Journal of Medicine The Seoul Journal of Medicine Vol. 22 No.4 (1981)
흰쥐 간조직 5'-Nucleotidase와 그 Isoenzyme에 관한 연구
On 5 nucleotidase and its isoenzymes in liver tissue of rat
- Issue Date
- 서울대학교 의과대학
- Seoul J Med, Vol.22 No.4, pp. 477-489
- 51-Nucleotidase is an intrisic membrane glycoprotein
which has been shown in a variety of cell types
to be an ectoenzyme_ It is widely used as a plasma
membrane marker in subcellular fractionation studies
and used in monitoring cell membrane changes during
It has been reported as that 5ιnucleotidase has
broad specificities for nucleoside 5' -monophosphate
Attempts have been made for such a reason to
resolve this enzyme into isoenzymes, but in most
cases evidence for isoenzymes were not obtained.
The present investigation was carried out to prepare
lipid free 5ιnucleotidase from rat liver and to study
its properties and isoenzymes with the following
1. Lipid-free 5ιnucleotidase obtained by Sephadex
G-200 gel filtration or DEAE-cellulose chromatography
from rat liver was resolved into two isoenzyme
bands by polyacrylamide gel disk electrophoresis.
2. 5'-Nucleotidase was purified about 45 folds
from rat liver homogenate with 42% yield by nbutanol
extraction and gel filtration, but the enzyme isoenzymepurified
by the treatment of n-butanol extraction
and DFAE'callulose chromatography showed the 11
folds of purification with 5% yield.
3. The pH optimum and KM values of the two
differently purified enzymes were identical; pH 7.5,
O. 33mM, respectively.
4. The enzymes showed different pattern in the
effect of EDTA; 5ιnucleotidase purified by gel
filtration method revealed strongly inhibited activity,
in contrast with that by DEAE-cellulose chromatography.
5. Effects of metal ions(Ca",Mg & Mn") on the
activity of 5ιnucleotidase by two procedures used
were similar to that by microsomal 5'-nucleotidase.
6. The effects of detergents (Triton X-IOO, sodium
deoxycholate and sodium dodecylsulfate) on the
activity of the enzymes were prominently different;
the enzyme by gel filtration showed increased activity,
while that by DEAE'cellulose chromatography, de'
The KM value , however was not altered, while