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Free access | 10.1172/JCI109370

Folate Distribution in Cultured Human Cells: STUDIES ON 5,10-CH2-H4PTEGLU REDUCTASE DEFICIENCY

David S. Rosenblatt, Bernard A. Cooper, and Sally Lue-Shing

MRC Genetics Group and Department of Pediatrics, McGill University, Montreal, Quebec H3H 1P3

Haematology Division, Royal Victoria Hospital, Montreal, Quebec H3H 1P3

Department of Physiology, McGill University, Montreal, Quebec H3H 1P3

Section of Genetics, Department of Pediatrics, Presbyterian-St. Luke's Medical Center, Rush Medical School, Chicago, Illinois 60612

Department of Pediatrics, Waisman Center on Mental Retardation and Human Development, University of Wisconsin, Madison, Wisconsin 53706

Department of Pediatrics, Tohoku University, School of Medicine, Sendai, Japan

Metabolic Unit, Basler Kinderspital, Basel, Switzerland

Find articles by Rosenblatt, D. in: PubMed | Google Scholar

MRC Genetics Group and Department of Pediatrics, McGill University, Montreal, Quebec H3H 1P3

Haematology Division, Royal Victoria Hospital, Montreal, Quebec H3H 1P3

Department of Physiology, McGill University, Montreal, Quebec H3H 1P3

Section of Genetics, Department of Pediatrics, Presbyterian-St. Luke's Medical Center, Rush Medical School, Chicago, Illinois 60612

Department of Pediatrics, Waisman Center on Mental Retardation and Human Development, University of Wisconsin, Madison, Wisconsin 53706

Department of Pediatrics, Tohoku University, School of Medicine, Sendai, Japan

Metabolic Unit, Basler Kinderspital, Basel, Switzerland

Find articles by Cooper, B. in: PubMed | Google Scholar

MRC Genetics Group and Department of Pediatrics, McGill University, Montreal, Quebec H3H 1P3

Haematology Division, Royal Victoria Hospital, Montreal, Quebec H3H 1P3

Department of Physiology, McGill University, Montreal, Quebec H3H 1P3

Section of Genetics, Department of Pediatrics, Presbyterian-St. Luke's Medical Center, Rush Medical School, Chicago, Illinois 60612

Department of Pediatrics, Waisman Center on Mental Retardation and Human Development, University of Wisconsin, Madison, Wisconsin 53706

Department of Pediatrics, Tohoku University, School of Medicine, Sendai, Japan

Metabolic Unit, Basler Kinderspital, Basel, Switzerland

Find articles by Lue-Shing, S. in: PubMed | Google Scholar

Published May 1, 1979 - More info

Published in Volume 63, Issue 5 on May 1, 1979
J Clin Invest. 1979;63(5):1019–1025. https://doi.org/10.1172/JCI109370.
© 1979 The American Society for Clinical Investigation
Published May 1, 1979 - Version history
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Abstract

We have studied the distribution of folate coenzyme forms in cultured human fibroblasts from control lines and from lines derived from nine patients representing all of the published reports of 5,10-CH2-H4PteGlu reductase deficiency. Based on mobility on DEAE-Sephadex and differential microbiological assay the major folate fractions in extracts of human fibroblasts were 5-CH3-H4PteGlu, 10-CHO-H4PteGlu, and 5-CHO-H4PteGlu with smaller fractions, which included 5-CH3-H2PteGlu, 10-CHO-PteGlu, and H4PteGlu. Evidence that the 5-CHO-H4PteGlu may have been derived from 5,10-CH=H4PteGlu during extraction is presented. In most of the mutant fibroblasts the absolute concentration of 5-CH3-H4PteGlu was lower than in control cells but the proportion of intracellular folate which was 5-CH3-H4PteGlu was strikingly lower in mutant cells when determined by chromatography or differential microbiological assay. In both control and mutant cells most of the 5-CH3-H4-PteGlu was polyglutamate. The proportion of intracellular folate which was polyglutamate was similar in control and mutant cells. A direct relationship was observed between the proportion of cellular folate which was 5-CH3-H4PteGlu, and both the clinical severity of this disorder and the residual enzyme activity indicating that the distribution of different folates may be an important control of intracellular folate metabolism. These studies indicate that 5,10-CH2-H4PteGlu reductase is the only significant intracellular pathway for the generation of 5-CH3-H4PteGlu, that the activity of this enzyme regulates the level of this folate in control and mutant cells under conditions of culture used here, that the majority of intracellular folate is in the polyglutamate form, and that the relative distribution of folates may control folate metabolism by interaction in the various folate reactions.

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