In vitro conversion of formate to serine: effect of tetrahydropteroylpolyglutamates and serine hydroxymethyltransferase on the rate of 10-formyltetrahydrofolate synthetase

Biochemistry ◽  
1989 ◽  
Vol 28 (24) ◽  
pp. 9430-9439 ◽  
Author(s):  
William B. Strong ◽  
Verne Schirch
Keyword(s):  
Serine Hydroxymethyltransferase ◽  
Formyltetrahydrofolate Synthetase

scholarly journals Regulation of C1 metabolism by l-methionine in Saccharomyces cerevisiae

1972 ◽  
Vol 130 (3) ◽  
pp. 773-783 ◽  
Author(s):  
K. L. Lor ◽  
E. A. Cossins
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Methylenetetrahydrofolate Reductase ◽  
Deae Cellulose ◽  
Serine Hydroxymethyltransferase ◽  
C1 Metabolism ◽  
Formyltetrahydrofolate Synthetase ◽  
Feeding Experiments ◽  
Cellulose Column ◽  
Homocysteine Methyltransferase

1. The concentrations of folate derivatives in aerobic cultures of Saccharomyces cerevisiae (A.T.C.C. 9763) were determined by microbiological assay employing Lactobacillus casei (A.T.C.C. 7469) and Pediococcus cerevisiae (A.T.C.C. 8081). Cells cultured in media lacking l-methionine contained higher concentrations of folate derivatives than cells grown in the same media supplemented with 2.5μmol of l-methionine/ml. The concentrations of highly conjugated derivatives were also decreased by supplementing the growth medium with l-methionine. 2. DEAE-cellulose column chromatography of extracts prepared from cells grown under these conditions revealed that the concentrations of methylated tetrahydrofolates were drastically decreased by the methionine supplement. Smaller decreases were also observed in the concentrations of formylated and unsubstituted derivatives. 3. The concentrations of four enzymes of C1 metabolism were compared after 6h of growth in the presence and in the absence of l-methionine (2.5μmol/ml). The specific activities of formyltetrahydrofolate synthetase, methylenetetrahydrofolate reductase and serine hydroxymethyltransferase were not altered by this treatment but that of 5-methyltetrahydrofolate–homocysteine methyltransferase was decreased by approx. 65% when l-methionine was supplied. The activities of 5-methyltetrahydrofolate–homocysteine methyltransferase, serine hydroxymethyltransferase and formyltetrahydrofolate synthetase were not appreciably altered by l-methionine in vitro. In contrast this amino acid was found to inhibit the activity of methylenetetrahydrofolate reductase. 4. Feeding experiments employing sodium [14C]formate indicated that cells grown in the presence of exogenous methionine, although having less ability to convert formate into methionine, readily incorporated 14C into serine and the adenosyl moiety of S-adenosylmethionine. 5. It is suggested that exogenous l-methionine controls C1 metabolism in Saccharomyces principally by regulation of methyl-group biogenesis within the folate pool.

scholarly journals Effects of thyroxine on the synthesis of folate coenzymes in rat liver

1970 ◽  
Vol 116 (2) ◽  
pp. 217-221 ◽  
Author(s):  
P. Pasquali ◽  
L. Landi ◽  
C. Bovina ◽  
M. Marchetti
Keyword(s):  
Rat Liver ◽  
Chronic Administration ◽  
Serine Hydroxymethyltransferase ◽  
Acute Administration ◽  
Formyltetrahydrofolate Synthetase ◽  
Derivatives Of

1. The effects of thyroidectomy and of ‘acute’ and ‘chronic’ administration of thyroxine on the synthesis of folate coenzymes were studied by determining the liver contents of folate active derivatives and the enzymic activities involved in their biosynthesis. The effect of thyroxine on the same enzymes in vitro was also studied. 2. In thyroidectomized rats the liver contents of folate coenzymes did not change except for a slight decrease in the contents of 5-formyltetrahydrofolate and tetrahydrofolate compared with those in control rats. 3. In the same animals serine hydroxymethyltransferase and formyltetrahydrofolate synthetase activities decreased markedly. 4. The ‘chronic’ administration of thyroxine to thyroidectomized rats caused more evident variations in the liver contents of folate coenzymes and in particular a decrease in the contents of 5-formyltetrahydrofolate, tetrahydrofolate, 5(or 10)-formyl derivatives of tetrahydropteroylpolyglutamate and of 5(or 10)-formyl derivatives of pteroylpolyglutamate. 5. The enzymic activities did not show significant variations. 6. The ‘acute’ administration of thyroxine caused changes in the liver contents of some folate derivatives such as 10-formyldihydrofolate, 10-formylfolate, tetrahydrofolate and the 10-formyl derivative of dihydropteroylpolyglutamate. In these animals also the enzymic activities were unchanged. 7. No effect of thyroxine on enzymic activities in vitro was observed.

scholarly journals The loss of SHMT2 mediates 5-fluorouracil chemoresistance in colorectal cancer by upregulating autophagy

Oncogene ◽  
2021 ◽  
Author(s):  
Jian Chen ◽  
Risi Na ◽  
Chao Xiao ◽  
Xiao Wang ◽  
Yupeng Wang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Patient Survival ◽  
Serine Hydroxymethyltransferase ◽  
First Line ◽  
Xenograft Models ◽  
Potential Opportunity ◽  
First Line Treatment ◽  
Novel Therapeutic

Abstract5-Fluorouracil (5-FU)-based chemotherapy is the first-line treatment for colorectal cancer (CRC) but is hampered by chemoresistance. Despite its impact on patient survival, the mechanism underlying chemoresistance against 5-FU remains poorly understood. Here, we identified serine hydroxymethyltransferase-2 (SHMT2) as a critical regulator of 5-FU chemoresistance in CRC. SHMT2 inhibits autophagy by binding cytosolic p53 instead of metabolism. SHMT2 prevents cytosolic p53 degradation by inhibiting the binding of p53 and HDM2. Under 5-FU treatment, SHMT2 depletion promotes autophagy and inhibits apoptosis. Autophagy inhibitors decrease low SHMT2-induced 5-FU resistance in vitro and in vivo. Finally, the lethality of 5-FU treatment to CRC cells was enhanced by treatment with the autophagy inhibitor chloroquine in patient-derived and CRC cell xenograft models. Taken together, our findings indicate that autophagy induced by low SHMT2 levels mediates 5-FU resistance in CRC. These results reveal the SHMT2–p53 interaction as a novel therapeutic target and provide a potential opportunity to reduce chemoresistance.

ACUTE EFFECTS OF TESTOSTERONE PROPIONATE ON FOLATE COENZYME SYNTHESIS IN THE RAT

1972 ◽  
Vol 54 (3) ◽  
pp. 457-464
Author(s):  
C. BOVINA ◽  
B. TOLOMELLI ◽  
C. ROVINETTI ◽  
M. MARCHETTI
Keyword(s):  
Folic Acid ◽  
Enzymic Activity ◽  
Male Rats ◽  
Formyltetrahydrofolate Synthetase ◽  
Folic Acid Metabolism ◽  
Testosterone Administration ◽  
Early Effects ◽  
Reduced Forms

SUMMARY The early effects of castration and testosterone administration on folic acid metabolism in liver and accessory sex organs were studied in adult male rats. After 3 days, castration did not produce significant variations in the folate coenzyme content of the liver, whereas in the prostate and seminal vesicles it only led to a fall of formylate forms. Testosterone administration did not cause significant changes in normal rats, but in castrated ones it generally raised formylate form levels and lowered the 5-methyltetrahydrofolate content. The capacity to convert folic acid into its activated forms in vivo decreased in castrated animals; after folic acid injection, they eliminated in urine more vitamin and fewer active forms than control rats. Furthermore, the liver content of the reduced forms was less in castrated animals than in intact ones. Testosterone administration partially adjusted this biochemical alteration. Studying in vitro the enzymic activities involved in folate coenzyme metabolism, it was found that the 10-formyltetrahydrofolate synthetase alteration was the more evident and constant one in castrated rats. This enzymic activity was much decreased in the organs studied. The other enzymes showed only a very slight increase in their activity. Testosterone administration to castrated rats not only prevented the fall but stimulated 10-formyltetrahydrofolate synthetase. The present data are discussed in relation to a direct or indirect effect of testosterone on enzymes involved in folate metabolism.

scholarly journals Interaction between glycine decarboxylase, serine hydroxymethyltransferase and tetrahydrofolate polyglutamates in pea leaf mitochondria

1994 ◽  
Vol 302 (1) ◽  
pp. 223-228 ◽  
Author(s):  
F Rebeille ◽  
M Neuburger ◽  
R Douce
Keyword(s):  
Dissociation Constants ◽  
Oxidative Degradation ◽  
Maximal Activity ◽  
Serine Hydroxymethyltransferase ◽  
In Vitro Experiments ◽  
Glycine Cleavage System ◽  
Kd Values ◽  
Bulk Medium ◽  
Glycine Cleavage

The aim of the present work was to further determine how the T-protein of the glycine-cleavage system and serine hydroxy-methyltransferase (SHMT), two folate-dependent enzymes from pea leaf mitochondria, interact through a common pool of tetrahydrofolate polyglutamates (H4PteGlun). It was observed that the binding affinity of tetrahydrofolate polyglutamates for these proteins continuously increased with increasing number of glutamates up to six residues. It was also established that, once bound to the proteins, tetrahydrofolate, a very O2-sensitive molecule, was protected from oxidative degradation. The dissociation constants (Kd) of H4PteGlu5, the most predominant form of polyglutamate in the mitochondria, were approximately 0.5 microM for both T-protein and SHMT, whereas the Kd values of CH2-H4PteGlu5 were higher, 2.7 and 7 microM respectively. In a matrix extract from pea leaf mitochondria, the maximal activity of the glycine-cleavage system was about 2.5 times higher than the maximal activity of SHMT. This resulted in a permanent disequilibrium of the SHMT-catalysed reaction which was therefore driven toward the production of serine and H4PteGlun, the thermodynamically unfavourable direction. Indeed, measurements of the steady-state ratio of CH2-H4PteGlun/H4PteGlun (n = 1 or n = 5) during the course of glycine oxidation demonstrated that the methylene form accounted for 65-80% of the folate pool. This indicates that, in our in vitro experiments, CH2-H4PteGlun with long polyglutamate chains accumulated in the bulk medium. This observation suggests that, in these in vitro experiments at least, there was no channelling of CH2-H4PteGlu5 between the T-protein and SHMT.

scholarly journals Screening and In Vitro Testing of Antifolate Inhibitors of Human Cytosolic Serine Hydroxymethyltransferase

ChemMedChem ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. 490-497 ◽  
Author(s):  
Alessandro Paiardini ◽  
Alessio Fiascarelli ◽  
Serena Rinaldo ◽  
Frederick Daidone ◽  
Giorgio Giardina ◽  
...  
Keyword(s):  
Serine Hydroxymethyltransferase ◽  
In Vitro Testing

In silico and in vitro validation of serine hydroxymethyltransferase as a chemotherapeutic target of the antifolate drug pemetrexed

2011 ◽  
Vol 46 (5) ◽  
pp. 1616-1621 ◽  
Author(s):  
Frederick Daidone ◽  
Rita Florio ◽  
Serena Rinaldo ◽  
Roberto Contestabile ◽  
Martino L. di Salvo ◽  
...  
Keyword(s):  
In Silico ◽  
Serine Hydroxymethyltransferase

scholarly journals Increased activity of renal glycine-cleavage-enzyme complex in metabolic acidosis

1985 ◽  
Vol 231 (2) ◽  
pp. 477-480 ◽  
Author(s):  
M Lowry ◽  
D E Hall ◽  
J T Brosnan
Keyword(s):  
Metabolic Acidosis ◽  
Serine Hydroxymethyltransferase ◽  
Enzyme Complex ◽  
Glycine Metabolism ◽  
Methylenetetrahydrofolate Dehydrogenase ◽  
Cleavage Enzyme ◽  
Combined Actions ◽  
Glycine Cleavage ◽  
Increased Activity

Glycine is metabolized in isolated renal cortical tubules to stochiometric qualities of ammonia, CO2 and serine by the combined actions of the glycine-cleavage-enzyme complex and serine hydroxymethyltransferase. The rate of renal glycine metabolism by this route is increased in tubules from acidotic rats, but is not affected in vitro by decreasing the incubation pH from 7.4 to 7.1. Metabolic acidosis caused an increase in the renal activity of the glycine-cleavage-enzyme complex, but there were no changes in the activity of serine hydroxymethyltransferase or of methylenetetrahydrofolate dehydrogenase. This enzymic adaptation permits increased ammoniagenesis from glycine during acidosis. The physiological implications are discussed.

scholarly journals Functional Identification of Serine Hydroxymethyltransferase as a Key Gene Involved in Lysostaphin Resistance and Virulence Potential of Staphylococcus aureus Strains

2020 ◽  
Vol 21 (23) ◽  
pp. 9135
Author(s):  
Nayab Batool ◽  
Kwan Soo Ko ◽  
Akhilesh Kumar Chaurasia ◽  
Kyeong Kyu Kim
Keyword(s):  
Staphylococcus Aureus ◽  
Serine Hydroxymethyltransferase ◽  
Multiple Drug ◽  
Host Immune System ◽  
Functional Identification ◽  
Resistance Development ◽  
Mammalian Cell Lines ◽  
Virulence Potential

Gaining an insight into the mechanism underlying antimicrobial-resistance development in Staphylococcus aureus is crucial for identifying effective antimicrobials. We isolated S. aureus sequence type 72 from a patient in whom the S. aureus infection was highly resistant to various antibiotics and lysostaphin, but no known resistance mechanisms could explain the mechanism of lysostaphin resistance. Genome-sequencing followed by subtractive and functional genomics revealed that serine hydroxymethyltransferase (glyA or shmT gene) plays a key role in lysostaphin resistance. Serine hydroxymethyltransferase (SHMT) is indispensable for the one-carbon metabolism of serine/glycine interconversion and is linked to folate metabolism. Functional studies revealed the involvement of SHMT in lysostaphin resistance, as ΔshmT was susceptible to the lysostaphin, while complementation of the knockout expressing shmT restored resistance against lysostaphin. In addition, the ΔshmT showed reduced virulence under in vitro (mammalian cell lines infection) and in vivo (wax-worm infection) models. The SHMT inhibitor, serine hydroxymethyltransferase inhibitor 1 (SHIN1), protected the 50% of the wax-worm infected with wild type S. aureus. These results suggest SHMT is relevant to the extreme susceptibility to lysostaphin and the host immune system. Thus, the current study established that SHMT plays a key role in lysostaphin resistance development and in determining the virulence potential of multiple drug-resistant S. aureus.

scholarly journals One-Carbon Metabolic Pathway Rewiring in Escherichia coli Reveals an Evolutionary Advantage of 10-Formyltetrahydrofolate Synthetase (Fhs) in Survival under Hypoxia

2014 ◽  
Vol 197 (4) ◽  
pp. 717-726 ◽  
Author(s):  
Shivjee Sah ◽  
Srinivas Aluri ◽  
Kervin Rex ◽  
Umesh Varshney
Keyword(s):  
Escherichia Coli ◽  
Anaerobic Bacteria ◽  
High Capacity ◽  
Natural Occurrence ◽  
Content Type ◽  
E Coli ◽  
Formyltetrahydrofolate Synthetase ◽  
Facultative Anaerobic Bacteria

In cells,N10-formyltetrahydrofolate (N10-fTHF) is required for formylation of eubacterial/organellar initiator tRNA and purine nucleotide biosynthesis. Biosynthesis ofN10-fTHF is catalyzed by 5,10-methylene-tetrahydrofolate dehydrogenase/cyclohydrolase (FolD) and/or 10-formyltetrahydrofolate synthetase (Fhs). All eubacteria possess FolD, but some possess both FolD and Fhs. However, the reasons for possessing Fhs in addition to FolD have remained unclear. We usedEscherichia coli, which naturally lacksfhs, as our model. We show that inE. coli, the essential function offolDcould be replaced byClostridium perfringensfhswhen it was provided on a medium-copy-number plasmid or integrated as a single-copy gene in the chromosome. Thefhs-supportedfolDdeletion (ΔfolD) strains grow well in a complex medium. However, these strains require purines and glycine as supplements for growth in M9 minimal medium. Thein vivolevels ofN10-fTHF in the ΔfolDstrain (supported by plasmid-bornefhs) were limiting despite the high capacity of the available Fhs to synthesizeN10-fTHFin vitro. Auxotrophy for purines could be alleviated by supplementing formate to the medium, and that for glycine was alleviated by engineering THF import into the cells. The ΔfolDstrain (harboringfhson the chromosome) showed a high NADP+-to-NADPH ratio and hypersensitivity to trimethoprim. The presence offhsinE. coliwas disadvantageous for its aerobic growth. However, under hypoxia,E. colistrains harboringfhsoutcompeted those lacking it. The computational analysis revealed a predominant natural occurrence offhsin anaerobic and facultative anaerobic bacteria.

Sign in / Sign up

Export Citation Format

Share Document