Search PubMed⌕ Search

Biomedical subjects

E L Stokstad

Publications and source records attributed to E L Stokstad.

At least 37 records · Page 2Linked to original sources
PubMed1982

Folate oligoglutamate:amino acid transpeptidase.

A new and previously unrecognized enzyme activity folate oligoglutamate:amino acid transpeptidase (folate transpeptidase), was detected in rat liver extracts. This activity involves the exchange of the terminal gamma-glutamyl residue (where PteGlu represents pteroylglutamic acid) of H4PteGlu-gamma-Glu, PteGlu-gamma-Glu, or p-aminobenzoyl-Glu-gamma-Glu for an amino acid such as [14C]glutamic acid, [14C]methionine, [14C]glutamine, or [14C]glycine. In the case of substrates PteGlu2 and [14C]Glu, the product is PteGlu-gamma-[14C]Glu. The enzyme responsible for catalyzing this activity was purified 39,000-fold by precipitation of inactive protein, fractionation on anionic and molecular sieve columns, and chromatography on diaminohexane-Sepharose. The enzyme functions maximally at pH 7.9, shows an absolute requirement for sulfhydryl reductants, and appears to have a molecular weight of 65,000. The function of folate transpeptidase is not known. A second, though much weaker, enzyme activity, tetrahydrofolic acid:amino acid ligase (folate ligase), co-purified with folate transpeptidase through all steps. Folate transpeptidase and the associated folate ligase activity were separated from folate polyglutamate synthetase during the initial steps of the purification procedure.

Acyltransferases↗
PubMed1982

Folate pentaglutamate and folate hexaglutamate mediated one-carbon metabolism.

The proposal that folate polyglutamate cofactors of different chain lengths function differently in metabolism was investigated. We identified the one-carbon units present in rat liver folates within each of the liver folate polyglutamate groups, the folate penta-, hexa-, and heptaglutamates. This identification revealed that at the pentaglutamate level, 5-methyl-H4folate was the major form, accounting for 18% of the total liver folates, with small amounts of H4folate. At the hexaglutamate level, 5-methyl-H4folate and H4folate were major forms, accounting for 17% and 22% of the liver folates, respectively. At the heptaglutamate level, 5-methyl-H4folate occurred in small amounts while H4folate predominated. The rats used here had been fed a diet low in methionine and were exposed to N2O gas. These results are qualitatively similar to those from rat brain [Brody, T., Shin, Y. S., & Stokstad, E. L. R. (1976) J. Neurochem. 27, 409--413] where 5-methyl-H4folate was a major fraction of the folate pentaglutamates but a minor, if detectable, part of the hexa- and heptaglutamates. The folates 5-methyl-H4PteGlu5-7 were metabolically active in the liver, as illustrated by the severe contraction in the amounts of these folates following an injection of methionine. This indicates that "folate-binding proteins" do not prevent 5-methyl-H4folates from entering into one-carbon metabolism.

Animals↗
PubMed1982

Nitrous oxide inactivates methionine synthetase in human liver.

Activity of methionine synthetase was measured in liver biopsies of seven patients who had received 50% to 70% nitrous oxide supplemented by a combination of a narcotic and/or barbiturate with or without a volatile anesthetic, and from seven patients who were anesthetized without nitrous oxide (control group). Methionine synthetase activity (+/- SE) averaged 219 +/- 28 nmol of methionine per hour per gran of liver in patients given nitrous oxide, and 414 +/- 29 in control patients. Inactivation of methionine synthetase progressively increased as the product of the concentration of nitrous oxide and the exposure time increased. These results in humans are similar to those in animals and suggest that inactivation of methionine synthetase may play a role in the development of the pathologic effects seen in patients and medical personnel after exposure to nitrous oxide.

5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗
PubMed1981

Availability of vitamin B6 and pantothenate in an average American diet in man.

The availability of vitamin B6 and pantothenate in an average American diet was assessed healthy male volunteers. The subjects received two types of diets, both nutritionally equivalent to the average American diet: period 1 (35 days), semipurified formula diet (low in both vitamins) with daily supplements of 1.1 mg pyridoxine and 8.2 mg pantothenate; period 2 (35 days), natural food sources, providing 2.3 mg vitamin B6 and 11.5 mg pantothenate/day; period 3 (21 days), formula diet, providing 2.7 mg pyridoxine and 8.2 mg pantothenate/day. Daily protein intake was 96 g throughout the study. Vitamins in food and urine samples were determined microbiologically and plasma pyridoxal phosphate by a tyrosine apodecarboxylase radioassay method. Compared to the availability of the pure vitamins as 100%, the availability of vitamin B6 ranged from 61 to 81% with a mean of 71% using plasma pyridoxal phosphate data, and ranged from 73 to 92% with a mean of 79% according to urinary vitamin B6 data. Availability of pantothenate ranged from 40 to 61% with a mean of 50%, according to urinary pantothenate data. The average American diet used in our study contained 1.7 and 5.8 mg/day of available vitamin B6 and pantothenate, respectively.

Adult↗
PubMed1981

Inactivation of methionine synthetase by nitrous oxide in mice.

To characterize the nitrous oxide-induced inhibition of the enzyme, methionine synthetase, we measured enzyme inactivation as a function of nitrous oxide concentration and exposure time. Mice exposed to 0.8 atm nitrous oxide exhibited more than a 50 per cent decrease in liver methionine synthetase activity within 30 min, and activity dropped to 5--25 per cent of the original value after a 4-hour exposure. Although 4-hour exposures to low nitrous oxide partial pressures (less than 0.05 atm) did not significantly alter methionine synthetase activity, higher concentrations of nitrous oxide caused a progressive inhibition over this time period. Continuous exposure to trace levels of nitrous oxide (approximately 1100 ppm) for eight to 22 days produced a small but significant reduction in liver and brain methionine synthetase activity. Methionine synthetase activity returned to control levels two to four days following inactivation. Other anesthetics (xenon, halothane, isoflurane, enflurane) did not produce inactivation.

5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗
PubMed1980

Metabolic responses of folic acid and related compounds to thyroxine in rats.

This study deals with the effects of thyroidectomy and feeding thyroid powder on histidine and folic acid metabolism. Normal rats maintained on a soy protein diet, low in methionine but supplemented with vitamin B-12, oxidize approx. 10% of an injected dose of [2-14C]histidine in 3 h and excrete low levels of formiminoglutamic acid. Addition of methionine increases histidine oxidation to approx. 20%. The feeding of thyroid powder or the injection of high levels of thyroxine decreases histidine oxidation and increases formiminoglutamic acid excretion. Surgical thyroidectomy at weaning increases histidine oxidation to approx. 45% and, thus, resembles the effect of methionine in promoting histidine oxidation and decreasing formiminoglutamic acid excretion. The feeding of methionine to the thyroidectomized animal further increases histidine oxidation to 65%. The distribution of folate forms in the liver was determined by column chromatography following administration of a dose of tritiated folic acid. In the normal animal, tetrahydrofolate accounts for 38% of the total folate present. The feeding of methionine increases this to 48%, which is consistent with the observed increase in histidine metabolism. Thyroidectomy increases the percentage of tetrahydrofolate to 63% and the feeding of methionine further increases it to 68%. The percentage of tetrahydrofolate relative to total folate is in proportion to the observed rate of histidine metabolism. The action of thyroidectomy in increasing histidine oxidation may be accounted for by its effect in increasing the proportion of tetrahydrofolate.

Animals↗
PubMed1980

Folate assay: a comparison of radioassay and microbiological methods.

Folate mono- and polyglutamate standards were assayed by a microbiological method and by four commercially available radioassay methods to evaluate the usefulness of radioassay techniques for the quantitation of naturally occurring folates. Folate monoglutamates exhibited different responses in the radioassay procedures, depending on the one-carbon constituent and oxidation state. Folate polyglutamates exhibited an increased response that varied depending on the folate concentration. The varied responses of folate mono- and polyglutamates in the radioassay procedures make this technique unsuitable for the determination of the mixture of folate derivatives that are normally encountered in biological extracts.

Biological Assay↗
PubMed1978

Absorption of mono- and polyglutamyl folates in zinc-depleted man.

The effect of experimental zinc depletion on the intestinal absorption of mono- and polyglutamyl folates was assessed in six healthy male subjects by measuring the increase in serum folate after an oral dose of pteroylmonoglutamate and pteroylheptaglutamate before and after initiation of a zinc-deficient diet. At the beginning of the study, no differences were detected in the absorption of pteroylmono- and heptaglutamate. After zinc depletion, the rise in serum folate after an oral dose of pteroylheptaglutamate was reduced by an average of 53% while pteroylmonoglutamate absorption appeared to be unchanged. These data suggest that zinc depletion results in a decreased intestinal hydrolysis of pteroylpolyglutamate and that intestinal conjugase in man is a zinc-dependent enzyme.

Adult↗
PubMed1977

Effect of methionine on the metabolism of formate and histidine by rats fed folate/vitamin B-12-methionine-deficient diet.

The metabolism of formate and histidine were compared in rats and in perfused livers of rats on diets deficient in vitamin B-12, methionine, and folic acid. Excretion of formate and formiminoglutamic acid, and the oxidation of [2-14C]histidine and [14C]formate to 14CO2 were measured. Liver folate levels decreased to 40% of normal on the vitamin B-12- and methionine-deficient diets but the rate of oxidation of histidine to CO2 in the whole animal decreased to 15% of normal. This indicated a reduction in the metabolic activity of the liver folates in vitamin B-12deficiency. Comparison of formate and histidine catabolism in folic acid deficiency showed that the oxidation of histine was decreased to 5% of normal but formate oxidation was decreased to only 30% of normal. This indicates that 25% of formate oxidation normally proceeds by a non-folate-dependent pathway.

Animals↗
PubMed1977

Identification of glutamate chain lengths of endogenous folylpoly-gamma-glutamates in rat tissues.

A simplified procedure for the determination of the glutamate chain lengths of endogenous tissue folate is described. Natural pteroylpoly-gamma-glutamates in tissue extracts, irrespective of their one-carbon moiety, were converted by a reductive cleavage procedure to a homologous series of p-aminobenzoylpoly-gamma-glutamates, differing only in glutamate chain length. Desalting and concentration of the extracts were achieved by absorbing the derivatives on active charcoal followed by their elution with ethanol:ammonia. Aminobenzoylpolyglutamates were separated by DEAD-cellulose chromatography and quantitated by a colorimetric procedure for primary aromatic amines. The major endogenous folates in rat liver and kidney were pteroylpentaglutamate derivatives with smaller amounts of pteroyltetra- and hexaglutamate.

Animals↗