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Literature summary for 1.5.1.20 extracted from

  • Froese, D.; Kopec, J.; Rembeza, E.; Bezerra, G.; Oberholzer, A.; Suormala, T.; Lutz, S.; Chalk, R.; Borkowska, O.; Baumgartner, M.; Yue, W.
    Structural basis for the regulation of human 5,10-methylenetetrahydrofolate reductase by phosphorylation and S-adenosylmethionine inhibition (2018), Nat. Commun., 9, 2261 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.5.1.20

Crystallization (Commentary)

Crystallization (Comment) Organism
purified enzyme in complex with FAD and SAH, X-ray diffraction structure determination and analysis at 2.5 A resolution Homo sapiens

Protein Variants

Protein Variants Comment Organism
additional information to examine the importance of the N-terminal serine-rich region to global protein phosphorylation, a recombinant HsMTHFR38-644 is produced, which removes the N-terminal 37 amino acids, including the entire serine-rich region as well as the poorly conserved C-terminal 12 amino acids predicted to be of high disorder. Purified HsMTHFR38-644 is not phosphorylated, determined by phosphorylation mapping, or native mass spectrometry, and treatment with CIP does not alter the protein molecular mass. Construction of truncated enzyme mutants, sMTHFR1-656 and HsMTHFR38-644. Dimeric mutant HsMTHFR38-644 is not phosphorylated Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
S-adenosylmethionine SAM, allosteric inhibition by the reaction product. MTHFR phosphorylation, identified on 11 N-terminal residues (16 in total), increases sensitivity to SAM binding and inhibition Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics for wild-type and mutant enzymes Homo sapiens
0.0224
-
 5,10-methylenetetrahydrofolate pH 6.6, 46°C, enzyme mutant HsMTHFR1-656 Homo sapiens
0.0235
-
 NADPH pH 6.6, 46°C, enzyme mutant HsMTHFR38-644 Homo sapiens
0.0255
-
 5,10-methylenetetrahydrofolate pH 6.6, 46°C, enzyme mutant HsMTHFR38-644 Homo sapiens
0.0355
-
 NADPH pH 6.6, 46°C, enzyme mutant HsMTHFR1-656 Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5,10-methylenetetrahydrofolate + NADPH + H+ Homo sapiens
-
 5-methyltetrahydrofolate + NADP+
-
 r

Organism

Organism UniProt Comment Textmining
Homo sapiens P42898
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein MTHFR phosphorylation, identified on 11 N-terminal residues (16 in total). MTHFR phosphorylation, identified on 11 N-terminal residues (16 in total), increases sensitivity to SAM binding and inhibition. The residues are Ser9, Ser10, Ser18, Ser20, Ser21, Ser23, Ser25, Ser26, Ser29, Ser30, and Thr34, located within the N-terminal serine-rich region, including the putative phosphorylation determining residue Thr34. Phosphorylation of three further amino acids in the catalytic domain (Tyr90, Thr94, and Ser103) and two in the regulatory domain (Ser394 and Thr451). Phosphorylation does not alter MTHFR kinetic parameters. Dimeric mutant HsMTHFR38-644 is not phosphorylated Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methylenetetrahydrofolate + NADPH + H+
-
 Homo sapiens 5-methyltetrahydrofolate + NADP+
-
 r

Subunits

Subunits Comment Organism
dimer an asymmetric MTHFR dimer with inter-domain flexibility. The extensive linker connects and interacts with both domains, the N-terminal catalytic domain (aa 40-337) consisting of an 8alpha/8beta TIM barrel, adorned with three extra alpha-helices, and the C-terminal regulatory domain (aa 363-644) making up a fold of two five-stranded beta-sheets arranged side-by-side in the core, flanked by a number of alpha-helices Homo sapiens

Synonyms

Synonyms Comment Organism
5,10-methylenetetrahydrofolate reductase
-
 Homo sapiens
HsMTHFR
-
 Homo sapiens
MTHFR
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
46
-
 assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
40.7
-
 NADPH pH 6.6, 46°C, enzyme mutant HsMTHFR1-656 Homo sapiens
40.7
-
 5,10-methylenetetrahydrofolate pH 6.6, 46°C, enzyme mutant HsMTHFR1-656 Homo sapiens
51.4
-
 NADPH pH 6.6, 46°C, enzyme mutant HsMTHFR38-644 Homo sapiens
51.4
-
 5,10-methylenetetrahydrofolate pH 6.6, 46°C, enzyme mutant HsMTHFR38-644 Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.6
-
 assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
additional information FAD is not required, NADPH is highly preferred before NADH Homo sapiens
NADP+
-
 Homo sapiens
NADPH
-
 Homo sapiens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0027
-
 S-adenosylmethionine pH 6.6, 46°C, enzyme mutant HsMTHFR1-656 Homo sapiens
0.021
-
 S-adenosylmethionine pH 6.6, 46°C, enzyme mutant HsMTHFR38-644 Homo sapiens

General Information

General Information Comment Organism
evolution human MTHFR is a 656 amino acid multi-domain protein, and the catalytic domain is conserved across evolution Homo sapiens
malfunction in accordance with its essential role, major and minor deficiencies of human MTHFR are the direct or indirect causes of human disease. Severe MTHFR deficiency is inherited in an autosomal recessive manner and is the most common inborn error of folate deficiency. Milder enzyme deficiencies, due to single nucleotide polymorphisms of the MTHFR gene, have been associated with various common disorders Homo sapiens
metabolism the folate and methionine cycles are crucial for biosynthesis of lipids, nucleotides and proteins, and production of the methyl donor S-adenosylmethionine (SAM). 5,10-methylenetetrahydrofolate reductase (MTHFR) represents a key regulatory connection between these cycles, generating 5-methyltetrahydrofolate for initiation of the methionine cycle, and undergoing allosteric inhibition by its end product SAM Homo sapiens
additional information human MTHFR reveals a unique architecture, appending the well-conserved catalytic TIM-barrel to a eukaryote-only SAM-binding domain. The latter domain of distinct fold provides the predominant interface for MTHFR homodimerization, positioning the N-terminal serine-rich phosphorylation region near the C-terminal SAM-binding domain. This explains how MTHFR phosphorylation, identified on 11 N-terminal residues (16 in total), increases sensitivity to SAM binding and inhibition. The 25-amino-acid inter-domain linker enables conformational plasticity and is proposed to be a key mediator of SAM regulation, molecular regulation of MTHFR, overview. The catalytic domain to form a beta8alpha8 (TIM) barrel, with residues critical for binding the cofactor FAD, the electron donor NADPH, and the product CH3-THF. The catalytic domain is sufficient for the entire catalytic cycle. Structure-function analysis, overview Homo sapiens
physiological function the folate and methionine cycles are crucial for biosynthesis of lipids, nucleotides and proteins, and production of the methyl donor S-adenosylmethionine (SAM). 5,10-methylenetetrahydrofolate reductase (MTHFR) represents a key regulatory connection between these cycles, generating 5-methyltetrahydrofolate for initiation of the methionine cycle, and undergoing allosteric inhibition by its end product SAM. Molecular regulation of MTHFR, overview. Phosphorylation does not alter MTHFR kinetic parameters. Phosphorylated MTHFR appeares to protect thermally unstable SAM from degradation to SAH, while the non-phosphorylated protein is unable to perform this function Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1146.5
-
 NADPH pH 6.6, 46°C, enzyme mutant HsMTHFR1-656 Homo sapiens
1817
-
 5,10-methylenetetrahydrofolate pH 6.6, 46°C, enzyme mutant HsMTHFR1-656 Homo sapiens
2015.7
-
 5,10-methylenetetrahydrofolate pH 6.6, 46°C, enzyme mutant HsMTHFR38-644 Homo sapiens
2187.2
-
 NADPH pH 6.6, 46°C, enzyme mutant HsMTHFR38-644 Homo sapiens