Adenosine deaminase from human erythrocytes: Purification and effects of adenosine analogs

Ram P. Agarwal, Stephen M. Sagar, Robert E. Parks

Research output: Contribution to journalArticle

179 Scopus citations

Abstract

Adenosine deaminase (adenosine aminohydrolase, EC 3.5.4.4) has been purified about 3000-fold from human erythrocytes. The molecular weight of the enzyme was estimated to be 33,000. With the partially purified erythrocytic adenosine deaminase, Km and Vmax values relative to adenosine were: adenosine, 25 μM, 100 per cent; formycin A, 1000 μM, 753-850 per cent; 8-aza-adenosine, 130 μM, 310 per cent; 6-chloropurine ribonuclcoside, 1000 μM, 91 per cent; 2,6-diaminopurine ribonucleoside, 74 μM, 91 per cent; 2'-deoxyadenosine. 7 μm, 60 per cent; xylosyladenine, 33 μm, 62 per cent; arabinosyi adenine, 100 μM, 47 per cent; 3'-deoxyadenosine (cordycepin), 41 μM, 100 per cent; 3'-amino3'-deoxyadenosine. 133 μM, 89 per cent: 4'-thioadenosine, 13 μM, 43 per cent; and 6-methylselenopurine ribonucleoside, 27 μM, 88 per cent. Apparent Kti values of reaction products and some adenosine analogs using adenosine as a substrate were as follows; inosine. 116 μM; 2'-deoxyinosine, 60 μM; guanosine, 140 μM; 2-fluoroadenosine, 60 μM; 2-fluorodeoxyadenosine. 19 μM; N6-methyladenosine, 17 μM; N1-methyladenosine, 275 μM; 6-thioguanosine, 92 μM; 6-thioinosine, 330 μM; 6-methylthioinosine, 270 μM; arabinosyl 6-thiopurine, 360 μM; and coformycin, 0.01 μM. Tubercidin (7-deaza-adenosine) and toyocamycin were devoid both of substrate and inhibitor activity. Also. N7-methylinosine, N7-methylguanosine and dipyridamole (Persantin®) did not inhibit the enzymic activity.

Original languageEnglish (US)
Pages (from-to)693-701
Number of pages9
JournalBiochemical Pharmacology
Volume24
Issue number6
DOIs
StatePublished - Mar 15 1975

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

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