Distribution of 5'-nucleotidase in muscle of some marine fishes.
A preliminary examination for the purification and characterization of 5'-nucleotidase of fish muscle was carried out and the following results were obtained. 1. The activities of 5'-nucleotidase in the muscles of marine vertebrates and invertebrates (total 11 species) were determined. The highest activity of 5'-nucleotidase was found in Blackrock fish Sebastes inermis, which was then used as a material for estimation of subcellular distribution and solubilization of the enzyme. 2. The 5'-nucleotidase of ordinary muscle of the fish Sebastes inermis was found in nuclear, microsomal and cytosolic fractions. About half of the total activity was found in the nuclear fraction, whereas the highest specific activity was observed in the microsomal fraction. 3. Complete solubilization of the enzyme was attained by using a high concentration of detergent such as Triton X-100, CHAPS, octylglucoside, octylthioglucoside and sodium deoxycholate, suggesting that the enzyme was tightly bound to the membrane. 4. Based on the results of solubility and stability tests, Triton X-100 seemed suitable for solubilizing 5'-nucleotidase from the membrane. 5. Microsomal 5'-nucleotidase was an Mg(2+)-activated enzyme, and no inactivation was observed up to 50 mM of Mg2+.
Marseno
D W
DW
Department of Food Science, Faculty of Applied Biological Science, Hiroshima University, Japan.
Hori
K
K
Miyazawa
K
K
D015720:5'-Nucleotidase / Q000378:metabolism*; D000818:Animals; D003902:Detergents; D005399:Fishes / Q000378:metabolism*; D015636:Magnesium Chloride / Q000494:pharmacology; D009132:Muscles / Q000201:enzymology*; D012995:Solubility
D003902:Detergents; D015636:Magnesium Chloride; D015720:5'-Nucleotidase