Soluble "high Km" 5'-nucleotidase activity in human T- and B-lymphoblasts: isolation and some properties.
1. Activity of "high Km" 5'-nucleotidase was investigated in the soluble fractions from cultured human T- and B-lymphoblasts. 2. Using gel filtration chromatography and 5'-AMP-Sepharose 4B affinity chromatography, it separated high Km 5'-nucleotidases from other two different soluble nucleoside 5'-phosphomonoesterase activities. 3. The molecular mass of the high Km enzymes from T- and B-lymphoblasts were 210 and 200 kDa, respectively. The optimum pH was at 6.5, and the Km values for IMP and AMP were 0.4 and 0.9 mM, respectively. 4. These properties of high Km 5'-nucleotidases were similar to those previously described from different tissues. These data indicate that soluble high Km 5'-nucleotidase coexists with "low Km" enzyme.
Madrid-Marina
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Department of Immunology and Rheumatology, Instituto Nacional de la Nutricion Salvador Zubiran, Mexico City, Mexico.
D015720:5'-Nucleotidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001402:B-Lymphocytes / Q000201:enzymology*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D008970:Molecular Weight; D012995:Solubility; D013601:T-Lymphocytes / Q000201:enzymology*
D015720:5'-Nucleotidase