Methylenetetrahydrofolate reductase

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methylene tetrahydrofolate reductase [NAD(P)H]
MTHFR active site.jpg
Ribbon diagram of the active site of E. coli MTHFR. The flavin cofactor (top) is shown interacting with the bound substrate NADH.[1]
Identifiers
EC number 1.5.1.20
CAS number 9028-69-7
Databases
IntEnz IntEnz view
BRENDA BRENDA entry
ExPASy NiceZyme view
KEGG KEGG entry
MetaCyc metabolic pathway
PRIAM profile
PDB structures RCSB PDB PDBe PDBsum
Gene Ontology AmiGO / EGO
Methylenetetrahydrofolate reductase (NAD(P)H)
Identifiers
Symbol MTHFR
External IDs OMIM607093 MGI106639 HomoloGene4349 GeneCards: MTHFR Gene
EC number 1.5.1.20
RNA expression pattern
PBB GE MTHFR 206800 at tn.png
PBB GE MTHFR 217071 s at tn.png
PBB GE MTHFR 217070 at tn.png
More reference expression data
Orthologs
Species Human Mouse
Entrez 4524 17769
Ensembl ENSG00000177000 ENSMUSG00000029009
UniProt P42898 Q9WU20
RefSeq (mRNA) NM_005957 NM_001161798
RefSeq (protein) NP_005948 NP_001155270
Location (UCSC) Chr 1:
11.85 – 11.87 Mb
Chr 4:
148.04 – 148.06 Mb
PubMed search [1] [2]

Methylenetetrahydrofolate reductase (MTHFR) is the rate-limiting enzyme in the methyl cycle, and it is encoded by the MTHFR gene.[2] Methylenetetrahydrofolate reductase catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, a cosubstrate for homocysteine remethylation to methionine. Genetic variation in this gene may influence susceptibility to occlusive vascular disease, neural tube defects, Alzheimer's disease and other forms of dementia, colon cancer, and acute leukemia, because mutations in this gene are associated with methylenetetrahydrofolate reductase deficiency.[3][4]

Biochemistry[edit]

In the rate-limiting step of the methyl cycle, MTHFR irreversibly reduces 5,10-methylenetetrahydrofolate (substrate) to 5-methyltetrahydrofolate (product).

MTHFR contains a bound flavin cofactor and uses NAD(P)H as the reducing agent.

Structure[edit]

"Secondary structure of MTHFR"
Secondary structure of MTHFR

Mammalian MTHFR is composed of an N-terminal catalytic domain and a C-terminal regulatory domain. MTHFR has at least two promoters and two isoforms (70 kDa and 77 kDa).[5]

Regulation[edit]

MTHFR activity may be inhibited by binding of dihydrofolate (DHF)[6] and S-adenosylmethionine (SAM, or AdoMet).[7] MTHFR can also be phosphorylated - this decreases its activity by ~20% and allows it to be more easily inhibited by SAM.[8]

Interactive pathway map[edit]

Click on genes, proteins and metabolites below to link to respective articles. [§ 1]

[[File:
FluoropyrimidineActivity_WP1601 go to article go to article go to article go to pathway article go to pathway article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to PubChem Compound go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to pathway article go to pathway article go to article go to article go to article go to article go to article go to WikiPathways go to article go to article go to article go to article go to article go to article go to article go to article go to article
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FluoropyrimidineActivity_WP1601 go to article go to article go to article go to pathway article go to pathway article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to PubChem Compound go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to article go to pathway article go to pathway article go to article go to article go to article go to article go to article go to WikiPathways go to article go to article go to article go to article go to article go to article go to article go to article go to article
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Fluorouracil (5-FU) Activity edit
  1. ^ The interactive pathway map can be edited at WikiPathways: "FluoropyrimidineActivity_WP1601". 

Genetics[edit]

The enzyme is coded by the gene with the symbol MTHFR on chromosome 1 location p36.3 in humans.[9] There are DNA sequence variants (genetic polymorphisms) associated with this gene. In 2000 a report brought the number of polymorphisms up to 24.[10] Two of the most investigated are C677T (rs1801133) and A1298C (rs1801131) single nucleotide polymorphisms (SNP).

C677T SNP (Ala222Val)[edit]

Main article: rs1801133

The MTHFR nucleotide at position 677 in the gene has two possibilities: C (cytosine) or T (thymine). C at position 677 (leading to an alanine at amino acid 222) is the normal allele. The 677T allele (leading to a valine substitution at amino acid 222) encodes a thermolabile enzyme with reduced activity.

Individuals with two copies of 677C (677CC) have the "normal" or "wildtype" genotype. 677TT individuals (homozygous) are said to have mild MTHFR deficiency. 677CT individuals (heterozygotes) are almost the same as normal individuals because the normal MTHFR can make up for the thermolabile MTHFR. About ten percent of the North American population are T-homozygous for this polymorphism. There is ethnic variability in the frequency of the T allele – frequency in Mediterranean/Hispanics is greater than the frequency in Caucasians which, in turn, is greater than in Africans/African-Americans.[11]

The degree of enzyme thermolability (assessed as residual activity after heat inactivation) is much greater in 677TT individuals (18-22%) compared with 677CT (56%) and 677CC (66-67%).[12] Individuals of 677TT are predisposed to mild hyperhomocysteinemia (high blood homocysteine levels), because they have less active MTHFR available to produce 5-methyltetrahydrofolate (which is used to decrease homocysteine). Low dietary intake of the vitamin folic acid can also cause mild hyperhomocysteinemia.

Low folate intake affects individuals with the 677TT genotype to a greater extent than those with the 677CC/CT genotypes. 677TT (but not 677CC/CT) individuals with lower plasma folate levels are at risk for elevated plasma homocysteine levels.[13] In studies of human recombinant MTHFR, the protein encoded by 677T loses its FAD cofactor three times faster than the wild-type protein.[14] 5-Methyl-THF slows the rate of FAD release in both the wild-type and mutant enzymes, although it is to a much greater extent in the mutant enzyme.[15] 677TT individuals are at an increased risk for certain leukemias[16] and colon cancer.[17]

Mutations in the MTHFR gene could be one of the factors leading to increased risk of developing schizophrenia.[18] Schizophrenic patients having the risk allele (T\T) show more deficiencies in executive function tasks.[19]

There is also a tentative link between MTHFR mutations and dementia. One study of an elderly Japanese population [20] found correlations between the MTHFR 677CT mutation, an Apo E polymorphism, and certain types of senile dementia. Other research has found that individuals with folate-related mutations can still have a functional deficiency even when blood levels of folate are within the normal range,[21] and recommended supplementation of methyltetrahydrofolate to potentially prevent and treat dementia (along with depression). A 2011 study[22] from China also found that the C677T SNP was associated with Alzheimer's disease in Asian populations (though not in Caucasians).

A1298C SNP (Glu429Ala)[edit]

At nucleotide 1298 of the MTHFR, there are two possibilities: A or C. 1298A (leading to a Glu at amino acid 429) is the most common while 1298C (leading to an Ala substitution at amino acid 429) is less common. 1298AA is the "normal" homozygous, 1298AC the heterozygous, and 1298CC the homozygous for the "variant". In studies of human recombinant MTHFR, the protein encoded by 1298C cannot be distinguished from 1298A in terms of activity, thermolability, FAD release, or the protective effect of 5-methyl-THF.[14] The C mutation does not appear to affect the MTHFR protein. It does not result in thermolabile MTHFR and does not appear to affect homocysteine levels. It does, however, affect the conversion of MTHF to BH4 (tetrahydrobiopterin), an important cofactor in the production of neurotransmitters, synthesis of nitric oxide, and detoxification of ammonia.

There has been some commentary on a 'reverse reaction' in which tetrahydrobiopterin (BH4) is produced when 5-methyltetrahydrofolate is converted back into methylenetetrahydrofolate. This however is not universally agreed upon. That reaction is thought to require 5-MTHF and SAMe.[citation needed] An alternative opinion is that 5-MTHF processes peroxynitrite, thereby preserving existing BH4, and that no such 'reverse reaction' occurs.

Detection of MTHFR Polymorphisms[edit]

A new triplex tetra-primer ARMS-PCR method was developed for the simultaneous detection of C677T and A1298C polymorphisms with the A66G MTRR polymorphism in a single PCR reaction (Lajin et al., 2012).

Severe MTHFR deficiency[edit]

Severe MTHFR deficiency is rare (about 50 cases worldwide) and caused by mutations resulting in 0-20% residual enzyme activity.[10] Patients exhibit developmental delay, motor and gait dysfunction, seizures, and neurological impairment and have extremely high levels of homocysteine in their plasma and urine as well as low to normal plasma methionine levels.

Reaction and metabolism[edit]

Schematic diagram of the reductive carbon-nitrogen bond cleavage (represented by wavy line) catalyzed by methylenetetrahydrofolate reductase.

The overall reaction catalyzed by MTHFR is illustrated on the right. The reaction uses an NAD(P)H hydride donor and an FAD cofactor. The E. coli enzyme has a strong preference for the NADH donor, whereas the mammalian enzyme is specific to NADPH.

MTHFR metabolism: folate cycle, methionine cycle, trans-sulfuration and hyperhomocysteinemia

As a drug target[edit]

Inhibitors of MTHFR and antisense knockdown of the expression of the enzyme have been proposed as treatments for cancer.[23]

References[edit]

  1. ^ PDB 1ZPT; Pejchal R, Sargeant R, Ludwig ML (2005). "Structures of NADH and CH3-H4folate complexes of Escherichia coli methylenetetrahydrofolate reductase reveal a spartan strategy for a ping-pong reaction". Biochemistry 44 (34): 11447–57. doi:10.1021/bi050533q. PMID 16114881. 
  2. ^ Goyette P, Sumner JS, Milos R, Duncan AM, Rosenblatt DS, Matthews RG, Rozen R (June 1994). "Human methylenetetrahydrofolate reductase: isolation of cDNA, mapping and mutation identification". Nat. Genet. 7 (2): 195–200. doi:10.1038/ng0694-195. PMID 7920641. 
  3. ^ "Entrez Gene: MTHFR methylenetetrahydrofolate reductase (NAD(P)H)". 
  4. ^ Födinger M, Hörl WH, Sunder-Plassmann G (2000). "Molecular biology of 5,10-methylenetetrahydrofolate reductase.". J Nephrol 13 (1): 20–33. PMID 10720211. 
  5. ^ Tran P, Leclerc D, Chan M, et al. (September 2002). "Multiple transcription start sites and alternative splicing in the methylenetetrahydrofolate reductase gene result in two enzyme isoforms". Mamm. Genome 13 (9): 483–92. doi:10.1007/s00335-002-2167-6. PMID 12370778. 
  6. ^ Matthews RG, Daubner SC (1982). "Modulation of methylenetetrahydrofolate reductase activity by S-adenosylmethionine and by dihydrofolate and its polyglutamate analogues". Adv. Enzyme Regul. 20: 123–31. doi:10.1016/0065-2571(82)90012-7. PMID 7051769. 
  7. ^ Jencks DA, Mathews RG (February 1987). "Allosteric inhibition of methylenetetrahydrofolate reductase by adenosylmethionine. Effects of adenosylmethionine and NADPH on the equilibrium between active and inactive forms of the enzyme and on the kinetics of approach to equilibrium". J. Biol. Chem. 262 (6): 2485–93. PMID 3818603. 
  8. ^ Yamada K, Strahler JR, Andrews PC, Matthews RG (July 2005). "Regulation of human methylenetetrahydrofolate reductase by phosphorylation". Proc. Natl. Acad. Sci. U.S.A. 102 (30): 10454–9. doi:10.1073/pnas.0504786102. PMC 1180802. PMID 16024724. 
  9. ^ Goyette P, Sumner JS, Milos R, et al. (August 1994). "Human methylenetetrahydrofolate reductase: isolation of cDNA mapping and mutation identification". Nat. Genet. 7 (4): 551. doi:10.1038/ng0894-551a. PMID 7951330. 
  10. ^ a b Sibani S, Christensen B, O'Ferrall E, Saadi I, Hiou-Tim F, Rosenblatt DS, Rozen R (2000). "Characterization of six novel mutations in the methylenetetrahydrofolate reductase (MTHFR) gene in patients with homocystinuria". Hum. Mutat. 15 (3): 280–7. doi:10.1002/(SICI)1098-1004(200003)15:3<280::AID-HUMU9>3.0.CO;2-I. PMID 10679944. 
  11. ^ Schneider JA, Rees DC, Liu YT, Clegg JB (May 1998). "Worldwide distribution of a common methylenetetrahydrofolate reductase mutation". Am. J. Hum. Genet. 62 (5): 1258–60. doi:10.1086/301836. PMC 1377093. PMID 9545406. 
  12. ^ Frosst P, Blom HJ, Milos R, Goyette P, Sheppard CA, Matthews RG, Boers GJ, den Heijer M, Kluijtmans LA, van den Heuvel LP et al. (May 1995). "A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase". Nat. Genet. 10 (1): 111–3. doi:10.1038/ng0595-111. PMID 7647779. 
  13. ^ Jacques PF, Bostom AG, Williams RR, Ellison RC, Eckfeldt JH, Rosenberg IH, Selhub J, Rozen R (January 1996). "Relation between folate status, a common mutation in methylenetetrahydrofolate reductase, and plasma homocysteine concentrations". Circulation 93 (1): 7–9. doi:10.1161/01.cir.93.1.7. PMID 8616944. 
  14. ^ a b Yamada K, Chen Z, Rozen R, Matthews RG (December 2001). "Effects of common polymorphisms on the properties of recombinant human methylenetetrahydrofolate reductase". Proc. Natl. Acad. Sci. U.S.A. 98 (26): 14853–8. doi:10.1073/pnas.261469998. PMC 64948. PMID 11742092. 
  15. ^ Schwahn B, Rozen R (2001). "Polymorphisms in the methylenetetrahydrofolate reductase gene: clinical consequences". Am J Pharmacogenomics 1 (3): 189–201. doi:10.2165/00129785-200101030-00004. PMID 12083967. 
  16. ^ Skibola CF, Smith MT, Kane E, Roman E, Rollinson S, Cartwright RA, Morgan G' (October 1999). "Polymorphisms in the methylenetetrahydrofolate reductase gene are associated with susceptibility to acute leukemia in adults". Proc. Natl. Acad. Sci. U.S.A. 96 (22): 12810–5. doi:10.1073/pnas.96.22.12810. PMC 23109. PMID 10536004. 
  17. ^ Ma J, Stampfer MJ, Giovannucci E, Artigas C, Hunter DJ, Fuchs C, Willett WC, Selhub J, Hennekens CH, Rozen R (15 March 1997). "Methylenetetrahydrofolate reductase polymorphism, dietary interactions, and risk of colorectal cancer". Cancer Res. 57 (6): 1098–102. PMID 9067278. 
  18. ^ "Meta-Analysis of All Published Schizophrenia-Association Studies (Case-Control Only) for rs1801133 (C677T) polymorphism, MTHFR gene". Schizophrenia Research Forum. Retrieved 2007-03-11. 
  19. ^ Roffman JL, Weiss AP, Deckersbach T, Freudenreich O, Henderson DC, Purcell S, Wong DH, Halsted CH, Goff DC (May 2007). "Effects of the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism on executive function in schizophrenia". Schizophr. Res. 92 (1–3): 181–8. doi:10.1016/j.schres.2007.01.003. PMID 17344026. 
  20. ^ http://www.ncbi.nlm.nih.gov/pubmed/10860300
  21. ^ http://www.ncbi.nlm.nih.gov/pubmed/17900207
  22. ^ http://www.ncbi.nlm.nih.gov/pubmed/21663380
  23. ^ Stankova J, Lawrance AK, Rozen R (2008). "Methylenetetrahydrofolate reductase (MTHFR): a novel target for cancer therapy". Curr Pharm Des 14 (11): 1143–50. doi:10.2174/138161208784246171. PMID 18473861. 

Further reading[edit]

  • Matthews RG (2003). "Methylenetetrahydrofolate reductase: a common human polymorphism and its biochemical implications". Chemical record 2 (1): 4–12. doi:10.1002/tcr.10006. PMID 11933257. 
  • Schwahn B, Rozen R (2002). "Polymorphisms in the methylenetetrahydrofolate reductase gene: clinical consequences". American journal of pharmacogenomics: genomics-related research in drug development and clinical practice 1 (3): 189–201. doi:10.2165/00129785-200101030-00004. PMID 12083967. 
  • Iqbal MP, Frossard PM (2003). "Methylene tetrahydrofolate reductase gene and coronary artery disease". JPMA. the Journal of the Pakistan Medical Association 53 (1): 33–6. PMID 12666851. 
  • Bailey LB (2003). "Folate, methyl-related nutrients, alcohol, and the MTHFR 677C-->T polymorphism affect cancer risk: intake recommendations". J. Nutr. 133 (11 Suppl 1): 3748S–3753S. PMID 14608109. 
  • Wiwanitkit V (2005). "Roles of methylenetetrahydrofolate reductase C677T polymorphism in repeated pregnancy loss". Clin. Appl. Thromb. Hemost. 11 (3): 343–5. doi:10.1177/107602960501100315. PMID 16015422. 
  • Muntjewerff JW, Kahn RS, Blom HJ, den Heijer M (2006). "Homocysteine, methylenetetrahydrofolate reductase and risk of schizophrenia: a meta-analysis". Mol. Psychiatry 11 (2): 143–9. doi:10.1038/sj.mp.4001746. PMID 16172608. 
  • Lewis SJ, Lawlor DA, Davey Smith G, et al. (2006). "The thermolabile variant of MTHFR is associated with depression in the British Women's Heart and Health Study and a meta-analysis". Mol. Psychiatry 11 (4): 352–60. doi:10.1038/sj.mp.4001790. PMID 16402130. 
  • Pereira TV, Rudnicki M, Pereira AC, et al. (2007). "5,10-Methylenetetrahydrofolate reductase polymorphisms and acute lymphoblastic leukemia risk: a meta-analysis". Cancer Epidemiol. Biomarkers Prev. 15 (10): 1956–63. doi:10.1158/1055-9965.EPI-06-0334. PMID 17035405. 
  • Leclerc D, Rozen R (2007). "[Molecular genetics of MTHFR: polymorphisms are not all benign]". Med Sci (Paris) 23 (3): 297–302. doi:10.1051/medsci/2007233297. PMID 17349292.