Adenylate kinase of human erythrocyte. Isolation and properties of the predominant inherited form.
Adenylate kinase exists in the human erythrocyte in a number of molecular forms with two allels at a single polymorphic locus coding for most of the enzyme forms. The predominant enzyme form, AK a, was purified to constant specific activity in excess of 3000 and appeared homogeneous by chromatography, electrophoresis, and ultracentrifugation. Sedimentation velocity and partial specific volume measurments of AK a yielded values of s20,w = 2.1 S and 0.722 cm-3per g. The molecular weight of the native enzyme was estimated to be 22,500 by sedimentation equilibrium and gel filtration analyses. The molecular weight of the denatured enzyme did not differ, indicating an absence of subunit structure in confirmation of genetic evidence of a single locus coding for the enzyme. The isolated enzyme demonstrated remarkable stability to denaturants (heat, guanidine HCl, urea) in the presence of appropriate stabilizing agents and could not be distinguished from rabbit muscle enzyme on this basis (as well as by a number of other kinetic and physicochemical parameters). The erythrocyte adenylate kinases have a common molecular size but differ in their charge properties. They demonstrate anomalous electrophoretic behavior, migrating anionic to hemoglobin in starch gel, yet exhibit isoelectric points considerably alkaline to hemoglobin (e.g. AK a, pI = 9.0) by isoelectric focusing.
D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002725:Chloroform; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004589:Electrophoresis, Disc; D004592:Electrophoresis, Starch Gel; D004912:Erythrocytes / Q000201:enzymology*; D005799:Genes, Dominant; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D000432:Methanol; D008970:Molecular Weight; D010641:Phenotype; D010770:Phosphotransferases / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D011817:Rabbits / Q000276:immunology; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014461:Ultracentrifugation
D002725:Chloroform; D010770:Phosphotransferases; D000432:Methanol