Characterization of the adenosine deaminase-adenosine deaminase complexing protein binding reaction.
Glutaraldehyde-fixed membranes from rabbit kidney cortex were used to characterize binding of monomeric adenosine deaminase to the adenosine deaminase complexing protein. With the use of bovine adenosine deaminase it was shown that enzyme binding is a saturable, high affinity process. The K value for binding of the bovine enzyme was 11 nM. Maximum enzyme binding and rate of binding to a constant amount of membrane did not vary significantly from pH 5.0 to 9.5. Metal ions, with the exception of Hg2+, sulfhydryl reagents, and other proteins had little or a slightly stimulatory effect on maximum binding. Mercuric ion inhibited binding. Using biotinylated bovine adenosine deaminase it was shown that purified rabbit, human, and monkey enzymes compete for binding sites on fixed membranes. The K values for the rabbit and human enzymes were 9 and 6 nM, respectively. Mouse or guinea pig adenosine deaminase did not bind to the membranes or compete with the biotinylated bovine enzyme for binding sites. The retention of characteristics required for binding by enzymes from rabbit, human, monkey, and calf tissues argues for biologic significance of the adenosine deaminase-complexing protein interaction. The basis for the apparent failure of rodent adenosine deaminase to bind to complexing protein remains to be determined.
Schrader
W P
WP
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201.
West
C A
CA
Miczek
A D
AD
Norton
E K
EK
D000243:Adenosine Deaminase / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002413:Cations, Divalent; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D006023:Glycoproteins / Q000378:metabolism*; D000882:Haplorhini; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism*; D007672:Kidney Cortex / Q000201:enzymology; D007700:Kinetics; D011485:Protein Binding; D011817:Rabbits; D013154:Spleen / Q000201:enzymology
D002413:Cations, Divalent; D006023:Glycoproteins; D007527:Isoenzymes; D000469:Alkaline Phosphatase; C489010:adenosine deaminase-complexing protein, Oryctolagus cuniculus; D000243:Adenosine Deaminase