Human Leukocyte Metabolism in Vitro. I. Incorporation of Adenine-8-C14 and Formate-C14 Into the Nucleic Acids of Leukemic Leukocytes

Scott, James

doi:10.1172/jci104468

Cited by 54 publications

(18 citation statements)

References 24 publications

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“…The rate of incorporation of ["C]formate into the normal leukocyte purines was found to be markedly lower, by about 10-fold, than that found in a previous study (10) for normal human cultured fibroblasts under similar conditions. Although this difference may in part reflect variation in endogenous formate pool size in these tissues, the low rate of activity of de novo purine synthesis in normal leukocytes, as gauged by formate incorporation, may be the explanation for the disagreement between various investigators concerning the presence of this pathway in these cells (2)(3)(4)(5).…”

Section: Discussionmentioning

confidence: 99%

“…Several investigators could not detect the complete pathway in normal leukocytes (2) or in lymphocytes (3), but demonstrated its presence in phytohemagglutinin-stimulated normal lymphocytes (3) and in leukemic cells (2,3). In contrast, other investigators did demonstrate de novo purine synthesis in normal leukocytes and found it to be accelerated in various hyperuricemic conditions such as primary gout, chronic renal failure, and secondary hyperuricemia (4,5).…”

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confidence: 99%

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De novo synthesis of purine nucleotides in human peripheral blood leukocytes. Excessive activity of the pathway in hypoxanthine-guanine phosphoribosyltransferase deficiency.

Brosh¹,

Boer²,

Kupfer³

et al. 1976

J. Clin. Invest.

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A B S T R A C T Human peripheral blood leukocytes were studied for the presence and the regulatory properties of the pathway of de novo synthesis of purine nucleotides. The cells were found to incorporate the labeled precursors formate and glycine into purines. The rate of [14C]-formate incorporation was decreased by several compounds known to inhibit purine synthesis by affecting the activity of glutamine phosphoribosylpyrophosphate (PRPP) amidotransferase, the first committed enzyme in the pathway, either through decreasing the availability of PRPP, a substrate for this enzyme, or through exerting inhibition on this enzyme.PRPP availability in the leukocyte was found to be limiting for purine synthesis. Increased PRPP availability resulting from activation of PRPP synthetase by increasing inorganic phosphate (Pi) concentration caused acceleration of purine synthesis. On the other hand, no clear-cut evidence was obtained for the availability of ribose-5-phosphate in the leukocyte being rate limiting at physiological extracellular Pi concentration for PRPP generation, and thus for purine synthesis. However, the addition of methylene blue, which accelerates the oxidative pentose shunt that produces ribose-5-phosphate, resulted in acceleration of PRPP gen- eration and of purine synthesis only when PRPP synthetase was largely activated at high Pi concentration. These results may be taken to suggest that ribose-5-phosphate availability is indeed not limiting for PRPP generation under physiological conditions. Purine synthesis de novo was accelerated more than 13-fold in the leukocytes of two gouty patients affected with partial deficiency of hypoxanthine-guanine phosphoribosyltransferase, but was normal in the leukocytes of an obligate heterozygote for this enzyme abnormality.The results demonstrate in peripheral human leukocytes the presence of the complete pathway of de novo synthesis of purine nucleotides and the manifestation in these cells of the biochemical consequences of hypoxanthine-guanine phosphoribosyltransferase deficiency, i.e., increased availability of PRPP and acceleration of purine synthesis de novo. The results indicate the usefulness of leukocytes as a model tissue for the study of purine metabolism in man.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

De novo synthesis of purine nucleotides in human peripheral blood leukocytes. Excessive activity of the pathway in hypoxanthine-guanine phosphoribosyltransferase deficiency.

Brosh¹,

Boer²,

Kupfer³

et al. 1976

J. Clin. Invest.

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“…Little work has been done on purine sources required for PHA stimulation, but almost all media used in the work on PHA effects on human lymphocytes, since the original description of the technique (18), have used media containing purines. It has been known for some time that lymphocytes have almost no capacity for synthesis of purine from simple precursors (23), but readily incorporate preformed purines into nucleic acids (23,24). Nevertheless, the most popular lymphocyte culture medias, the RMPl series, do not have a purine source (29).…”

Section: Discussionmentioning

confidence: 99%

Purine Dysfunction in Cells from Patients with Adenosine Deaminase Deficiency

Benke

Dittmar

1976

Pediatric Research

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ExtractConversion of adenosine to inosine is decreased in adenosine deaminase (ADA)-deficient fibroblasts at all concentrations of adenosine tested. Adenosine is not differentially toxic to ADA-deficient fibroblasts except at very high ( 5 x 10-'-1 x 10-3M) adenosine levels. Conversion of [14C]adenosine to GTP is not decreased in ADA-deficient cells compared with control cell strains. Adenosine conversion to ATP is the same as that in mutant cells except at high nonphysiologic concentrations, at which it is slightly decreased in ADA-deficient fibroblasts. This effect is probably not related to the biochemical pathology of ADA-deficient lymphocytes in vivo.

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“…Burkitt lymphoma cells 8 were maintained in tissue culture. Cells were grown in RPM1 1640 medium containing 20% fetal calf serum and supplemented with 200 ,umoles l-glutamine, 10,000 units penicillin, 10,000 ug streptomycin, and 25 ,ug Fungizone per 100 ml. The medium and all ingredients were obtained from Grand Island Biological Co., Berkeley, Calif.…”

Section: Methodsmentioning

confidence: 99%

De Novo purine biosynthesis by two pathways in Burkitt lymphoma cells and in human spleen

Reem¹

1972

J. Clin. Invest.

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Human Leukocyte Metabolism in Vitro. I. Incorporation of Adenine-8-C14 and Formate-C14 Into the Nucleic Acids of Leukemic Leukocytes

Cited by 54 publications

References 24 publications

De novo synthesis of purine nucleotides in human peripheral blood leukocytes. Excessive activity of the pathway in hypoxanthine-guanine phosphoribosyltransferase deficiency.

De novo synthesis of purine nucleotides in human peripheral blood leukocytes. Excessive activity of the pathway in hypoxanthine-guanine phosphoribosyltransferase deficiency.

Purine Dysfunction in Cells from Patients with Adenosine Deaminase Deficiency

De Novo purine biosynthesis by two pathways in Burkitt lymphoma cells and in human spleen

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