The presence and activity in normal and regenerating rat liver postmicrosomal supernatant fraction of an enzyme with properties similar to those of membrane-bound 5'-nucleotidase.
An alkaline 5'-nucleotidase with properties similar to those of membrane-bound 5'-nucleotidase was recovered in soluble form in the postmicrosomal supernatant fraction (cytosol) of rat liver. The enzyme seems to constitute a quantitatively distinct fraction, since the activity in postmicrosomal supernatants was increased by a further 10% by additional homogenization of livers. Lysosomal acid phosphatase activity increased similarly, whereas other membrane-bound marker enzymes alkaline phosphatase, phosphodiesterase I and glucose-6-phosphatase showed no increase when homogenization of liver tissue was continued. Gel-permeation chromatography and pH-dependence studies indicated that enzyme activity in the supernatant fraction with 0.3 mM-UMP or -AMP as substrate at pH 8.1 was about 85 or 100% specific respectively. In regenerating liver the enzyme recovered in soluble form showed decreased specific activity, in contrast with alkaline phosphatase measured for comparison. The nucleotidase activity per mg of cytosolic protein was 2.1 nmol/min with AMP as substrate. The total activity measured in the postmicrosomal supernatant was 1.5% of the homogenate activity measured in the presence of detergent.
D009597:4-Nitrophenylphosphatase / Q000378:metabolism; D015720:5'-Nucleotidase; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002474:Cell-Free System; D002850:Chromatography, Gel; D008099:Liver / Q000201:enzymology*; D008115:Liver Regeneration; D008297:Male; D009708:Nucleotidases / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D009711:Nucleotides; D011506:Proteins; D009708:Nucleotidases; D009597:4-Nitrophenylphosphatase; D015720:5'-Nucleotidase