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Molecular Pharmacology

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Research ArticleArticle

Inhibition by Nitrobenzylthioinosine of Adenosine Uptake by Asynchronous HeLa Cells

ALAN R. P. PATERSON, LORI R. BABB, JOHN H. PARAN and CAROL E. CASS
Molecular Pharmacology November 1977, 13 (6) 1147-1158;
ALAN R. P. PATERSON
Cancer Research Unit (McEachern Laboratory) and Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7
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LORI R. BABB
Cancer Research Unit (McEachern Laboratory) and Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7
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JOHN H. PARAN
Cancer Research Unit (McEachern Laboratory) and Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7
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CAROL E. CASS
Cancer Research Unit (McEachern Laboratory) and Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7
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Abstract

Initial rates of adenosine uptake by HeLa cell monolayers were measured by a replicate culture technique. Under the assay conditions, the cellular content of adenosine was almost entirely in the form of metabolites, primarily ATP, ADP, and AMP, and thus the time course of adenosine uptake was essentially that of the formation of the adenosine phosphates; both time courses were linear and could be extrapolated through zero time. Adenosine uptake was mediated by a mechanism that appeared to be distinct from the thymidine, uridine, and guanosine uptake mechanisms. In the presence of nitrobenzylthioinosine (NBMPR), mediated entry of adenosine was eliminated and a nonsaturable component of uptake, evidently simple diffusion, was apparent. In the presence of partially inhibitory concentrations of NBMPR, the apparent Km values of the uptake process were increased and Vmax values were unchanged. In a structure-activity study, comparisons were made of inhibition of adenosine uptake by graded concentrations of various nucleoside derivatives related to NBMPR. Inhibitors more potent than NBMPR were identified, and it was apparent that the 2'-hydroxyl group of NBMPR and related compounds was not involved in the cell-inhibitor interaction. NBMPR and its 5'-monophosphate were comparable inhibitors of adenosine uptake.

ACKNOWLEDGMENTS We acknowledge with gratitude the generous provision of compounds for testing by the following: T. Fujii, J. P. Miller, R. A. Long, M. J. Robins, H. J. Schaeffer, L. R. Townsend, H. Vorbrüggen, and Developmental Therapeutics Program, National Cancer Institute, Bethesda, Md.

  • Copyright © 1977 by Academic Press, Inc.

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Molecular Pharmacology
Vol. 13, Issue 6
1 Nov 1977
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Research ArticleArticle

Inhibition by Nitrobenzylthioinosine of Adenosine Uptake by Asynchronous HeLa Cells

ALAN R. P. PATERSON, LORI R. BABB, JOHN H. PARAN and CAROL E. CASS
Molecular Pharmacology November 1, 1977, 13 (6) 1147-1158;

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Research ArticleArticle

Inhibition by Nitrobenzylthioinosine of Adenosine Uptake by Asynchronous HeLa Cells

ALAN R. P. PATERSON, LORI R. BABB, JOHN H. PARAN and CAROL E. CASS
Molecular Pharmacology November 1, 1977, 13 (6) 1147-1158;
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