Dynamic inter-domain transformations mediate the allosteric regulation of human 5, 10-methylenetetrahydrofolate reductase.


Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
15 Apr 2024
Historique:
received: 11 09 2023
accepted: 22 03 2024
medline: 16 4 2024
pubmed: 16 4 2024
entrez: 15 4 2024
Statut: epublish

Résumé

5,10-methylenetetrahydrofolate reductase (MTHFR) commits folate-derived one-carbon units to generate the methyl-donor S-adenosyl-L-methionine (SAM). Eukaryotic MTHFR appends to the well-conserved catalytic domain (CD) a unique regulatory domain (RD) that confers feedback inhibition by SAM. Here we determine the cryo-electron microscopy structures of human MTHFR bound to SAM and its demethylated product S-adenosyl-L-homocysteine (SAH). In the active state, with the RD bound to a single SAH, the CD is flexible and exposes its active site for catalysis. However, in the inhibited state the RD pocket is remodelled, exposing a second SAM-binding site that was previously occluded. Dual-SAM bound MTHFR demonstrates a substantially rearranged inter-domain linker that reorients the CD, inserts a loop into the active site, positions Tyr404 to bind the cofactor FAD, and blocks substrate access. Our data therefore explain the long-distance regulatory mechanism of MTHFR inhibition, underpinned by the transition between dual-SAM and single-SAH binding in response to cellular methylation status.

Identifiants

pubmed: 38622112
doi: 10.1038/s41467-024-47174-y
pii: 10.1038/s41467-024-47174-y
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

3248

Subventions

Organisme : Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Swiss National Science Foundation)
ID : 310030_192505

Informations de copyright

© 2024. The Author(s).

Références

Petrossian, T. C. & Clarke, S. G. Uncovering the human methyltransferasome. Mol. Cell Proteom. 10, M110.000976 (2011).
doi: 10.1074/mcp.M110.000976
Lu, S. C. S-Adenosylmethionine. Int J. Biochem. Cell Biol. 32, 391–395 (2000).
pubmed: 10762064 doi: 10.1016/S1357-2725(99)00139-9
Igari, S. et al. Properties and crystal structure of methylenetetrahydrofolate reductase from Thermus thermophilus HB8. PLoS One 6, e23716 (2011).
pubmed: 21858212 pmcid: 3156243 doi: 10.1371/journal.pone.0023716
Liew, S. C. & Gupta, E. D. Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism: Epidemiology, metabolism and the associated diseases. Eur. J. Med. Genet. 58, 1–10 (2015).
pubmed: 25449138 doi: 10.1016/j.ejmg.2014.10.004
Izmirli, M. A literature review of MTHFR (C677T and A1298C polymorphisms) and cancer risk. Mol. Biol. Rep. 40, 625–637 (2012).
pubmed: 23076526 doi: 10.1007/s11033-012-2101-2
Frosst, P. et al. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat. Genet. 10, 111–113 (1995).
pubmed: 7647779 doi: 10.1038/ng0595-111
Raghubeer, S., Matsha, T. E., Mccaddon, A. & Miller, J. W. Methylenetetrahydrofolate (MTHFR), the One-Carbon Cycle, and Cardiovascular Risks. Nutrients 13, 4562 (2021).
pubmed: 34960114 pmcid: 8703276 doi: 10.3390/nu13124562
Parle-McDermott, A. et al. Analysis of the MTHFR 1298A→C and 677C→T polymorphisms as risk factors for neural tube defects. J. Hum. Genet. 48, 190–193 (2003).
pubmed: 12730722 doi: 10.1007/s10038-003-0008-4
Kirke, P. N. et al. Impact of the MTHFR C677T polymorphism on risk of neural tube defects: case-control study. BMJ 328, 1535–1536 (2004).
pubmed: 15155469 pmcid: 437144 doi: 10.1136/bmj.38036.646030.EE
Froese, D. S. et al. Mutation update and review of severe methylenetetrahydrofolate reductase deficiency. Hum. Mutat. 37, 427–438 (2016).
pubmed: 26872964 doi: 10.1002/humu.22970
Kutzbach, C. & Stokstad, E. L. R. Mammalian methylenetetrahydrofolate reductase. Partial purification, properties, and inhibition by S-adenosylmethionine. Biochim. Biophys. Acta 250, 459–477 (1971).
pubmed: 4399897 doi: 10.1016/0005-2744(71)90247-6
Sumner, J., Jencks, D. A., Khani, S. & Matthews, R. G. Photoaffinity labeling of methylenetetrahydrofolate reductase with 8-azido-S-adenosylmethionine. J. Biol. Chem. 261, 7697–7700 (1986).
pubmed: 3754872 doi: 10.1016/S0021-9258(19)57456-7
Daubner, S. C. & Matthews, R. G. Purification and properties of methylenetetrahydrofolate reductase from pig liver. J. Biol. Chem. 257, 140–145 (1982).
pubmed: 6975779 doi: 10.1016/S0021-9258(19)68337-7
Yamada, K., Chen, Z., Rozen, R. & Matthews, R. G. Effects of common polymorphisms on the properties of recombinant human methylenetetrahydrofolate reductase. Proc. Natl Acad. Sci. USA 98, 14853–14858 (2001).
pubmed: 11742092 pmcid: 64948 doi: 10.1073/pnas.261469998
Yamada, K., Strahler, J. R., Andrews, P. C. & Matthews, R. G. Regulation of human methylenetetrahydrofolate reductase by phosphorylation. Proc. Natl Acad. Sci. USA 102, 10454–10459 (2005).
pubmed: 16024724 pmcid: 1180802 doi: 10.1073/pnas.0504786102
Froese, D. S. et al. Structural basis for the regulation of human 5,10-methylenetetrahydrofolate reductase by phosphorylation and S-adenosylmethionine inhibition. Nat. Commun. 9, 1–13 (2018).
doi: 10.1038/s41467-018-04735-2
Matthews, R. G., Vanon, M. A., Hainfeld, J. F. & Wall, J. Methylenetetrahydrofolate reductase. Evidence for spatially distinct subunit domains obtained by scanning transmission electron microscopy and limited proteolysis. J. Biol. Chem. 259, 11647–11650 (1984).
pubmed: 6384210 doi: 10.1016/S0021-9258(20)71253-6
Bezerra, G. A. et al. Identification of small molecule allosteric modulators of 5,10-methylenetetrahydrofolate reductase (MTHFR) by targeting its unique regulatory domain. Biochimie 183, 100–107 (2021).
pubmed: 33476699 pmcid: 8040968 doi: 10.1016/j.biochi.2021.01.007
Guenther, B. D. et al. The structure and properties of methylenetetrahydrofolate reductase from Escherichia coli suggest how folate ameliorates human hyperhomocysteinemia. Nat. Struct. Biol. 6, 359–365 (1999).
pubmed: 10201405 doi: 10.1038/7594
Pejchal, R. et al. Structural perturbations in the Ala → Val polymorphism of methylenetetrahydrofolate reductase: How binding of folates may protect against inactivation. Biochemistry 45, 4808–4818 (2006).
pubmed: 16605249 doi: 10.1021/bi052294c
Gana, R., Rao, S., Huang, H., Wu, C. & Vasudevan, S. Structural and functional studies of S-adenosyl-L-methionine binding proteins: a ligand-centric approach. BMC Struct. Biol. 13, 6 (2013).
pubmed: 23617634 pmcid: 3662625 doi: 10.1186/1472-6807-13-6
Lee, M. N. et al. A Functional Role for the Conformationally Mobile Phenylalanine 223 in the Reaction of Methylenetetrahydrofolate Reductase from E. coli. Biochemistry 48, 7673 (2009).
pubmed: 19610625 doi: 10.1021/bi9007325
Greber, B. J., Remis, J., Ali, S. & Nogales, E. 2.5 Å-resolution structure of human CDK-activating kinase bound to the clinical inhibitor ICEC0942. Biophys. J. 120, 677–686 (2021).
pubmed: 33476598 pmcid: 7896097 doi: 10.1016/j.bpj.2020.12.030
Pejchal, R., Sargeant, R. & Ludwig, M. L. Structures of NADH and CH3-H4folate complexes of Escherichia coli methylenetetrahydrofolate reductase reveal a Spartan strategy for a ping-pong reaction. Biochemistry 44, 11447–11457 (2005).
pubmed: 16114881 doi: 10.1021/bi050533q
Trimmer, E. E., Ballou, D. P. & Matthews, R. G. Methylenetetrahydrofolate reductase from escherichia coli: Elucidation of the kinetic mechanism by steady-state and rapid-reaction studies. Biochemistry 40, 6205–6215 (2001).
pubmed: 11371181 doi: 10.1021/bi002789w
Sheppard, C. A., Trimmer, E. E. & Matthews, R. G. Purification and properties of NADH-dependent 5,10- methylenetetrahydrofolate reductase (MetF) from Escherichia coli. J. Bacteriol. 181, 718–725 (1999).
pubmed: 9922232 pmcid: 93435 doi: 10.1128/JB.181.3.718-725.1999
Burda, P. et al. Insights into Severe 5,10-Methylenetetrahydrofolate Reductase Deficiency: Molecular Genetic and Enzymatic Characterization of 76 Patients. Hum. Mutat. 36, 611–621 (2015).
pubmed: 25736335 doi: 10.1002/humu.22779
Burda, P. et al. Functional characterization of missense mutations in severe methylenetetrahydrofolate reductase deficiency using a human expression system. J. Inherit. Metab. Dis. 40, 297–306 (2017).
pubmed: 27743313 doi: 10.1007/s10545-016-9987-0
Bhatia, M. et al. Allosteric inhibition of MTHFR prevents futile SAM cycling and maintains nucleotide pools in one-carbon metabolism. J. Biol. Chem. 295, 16037–16057 (2020).
pubmed: 32934008 pmcid: 7681022 doi: 10.1074/jbc.RA120.015129
McCorvie, T. J. et al. Inter-domain Communication of Human Cystathionine β-Synthase. J. Biol. Chem. 289, 36018–36030 (2014).
pubmed: 25336647 pmcid: 4276868 doi: 10.1074/jbc.M114.610782
Patel, D., Kopec, J., Fitzpatrick, F., Mccorvie, T. J. & Yue, W. W. Structural basis for ligand-dependent dimerization of phenylalanine hydroxylase regulatory domain. Sci. Rep. 6, 1–10 (2016).
doi: 10.1038/srep23748
Jiao, W., Lang, E. J., Bai, Y., Fan, Y. & Parker, E. J. Diverse allosteric componentry and mechanisms control entry into aromatic metabolite biosynthesis. Curr. Opin. Struct. Biol. 65, 159–167 (2020).
pubmed: 32739636 doi: 10.1016/j.sbi.2020.06.015
Jumper, J. et al. Highly accurate protein structure prediction with AlphaFold. Nature 596, 583–589 (2021).
pubmed: 34265844 pmcid: 8371605 doi: 10.1038/s41586-021-03819-2
Varadi, M. et al. AlphaFold Protein Structure Database: massively expanding the structural coverage of protein-sequence space with high-accuracy models. Nucleic Acids Res. 50, D439–D444 (2022).
pubmed: 34791371 doi: 10.1093/nar/gkab1061
Goyette, P. et al. Human methylenetetrahydrofolate reductase: isolation of cDNA, mapping and mutation identification. Nat. Genet. 7, 195–200 (1994).
pubmed: 7920641 doi: 10.1038/ng0694-195
Layer, G., Moser, J., Heinz, D. W., Jahn, D. & Schubert, W. D. Crystal structure of coproporphyrinogen III oxidase reveals cofactor geometry of Radical SAM enzymes. EMBO J. 22, 6214–6224 (2003).
pubmed: 14633981 pmcid: 291839 doi: 10.1093/emboj/cdg598
Mas-Droux, C., Biou, V. & Dumas, R. Allosteric threonine synthase: Reorganization of the pyridoxal phosphate site upon asymmetric activation through S-adenosylmethionine binding to a novel site. J. Biol. Chem. 281, 5188–5196 (2006).
pubmed: 16319072 doi: 10.1074/jbc.M509798200
Jencks, D. A. & Mathews, R. G. 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, 2485–2493 (1987).
pubmed: 3818603 doi: 10.1016/S0021-9258(18)61530-3
LeGros, H. L., Halim, A. B., Geller, A. M. & Kotb, M. Cloning, expression, and functional characterization of the β regulatory subunit of human methionine adenosyltransferase (MAT II). J. Biol. Chem. 275, 2359–2366 (2000).
pubmed: 10644686 doi: 10.1074/jbc.275.4.2359
Weile, J. et al. Shifting landscapes of human MTHFR missense-variant effects. Am. J. Hum. Genet. 108, 1283–1300 (2021).
doi: 10.1016/j.ajhg.2021.05.009 pubmed: 34861178 pmcid: 8715197
Suormala, T., Gamse, G. & Fowler, B. 5,10-Methylenetetrahydrofolate Reductase (MTHFR) Assay in the Forward Direction: Residual Activity in MTHFR Deficiency. Clin. Chem. 48, 835–843 (2002).
pubmed: 12028998 doi: 10.1093/clinchem/48.6.835
Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat. Methods 14, 290–296 (2017).
pubmed: 28165473 doi: 10.1038/nmeth.4169
Huiskonen, J. T. Image processing for cryogenic transmission electron microscopy of symmetry-mismatched complexes. Biosci. Rep. 38, 20170203 (2018).
doi: 10.1042/BSR20170203
Pettersen, E. F. et al. UCSF ChimeraX: Structure visualization for researchers, educators, and developers. Protein Sci. 30, 70–82 (2021).
pubmed: 32881101 doi: 10.1002/pro.3943
Kidmose, R. T. et al. Namdinator - automatic molecular dynamics flexible fitting of structural models into cryo-EM and crystallography experimental maps. IUCrJ 6, 526–531 (2019).
pubmed: 31316797 pmcid: 6608625 doi: 10.1107/S2052252519007619
Waterhouse, A. et al. SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 46, W296–W303 (2018).
pubmed: 29788355 pmcid: 6030848 doi: 10.1093/nar/gky427
Liebschner, D. et al. Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr. D. Struct. Biol. 75, 861–877 (2019).
pubmed: 31588918 pmcid: 6778852 doi: 10.1107/S2059798319011471
Croll, T. I. ISOLDE: a physically realistic environment for model building into low-resolution electron-density maps. Acta Crystallogr. D. Struct. Biol. 74, 519–530 (2018).
pubmed: 29872003 pmcid: 6096486 doi: 10.1107/S2059798318002425
Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D. Biol. Crystallogr. 60, 2126–2132 (2004).
pubmed: 15572765 doi: 10.1107/S0907444904019158
Afonine, P. V. et al. Real-space refinement in PHENIX for cryo-EM and crystallography. Acta Crystallogr. D. Struct. Biol. 74, 531–544 (2018).
pubmed: 29872004 pmcid: 6096492 doi: 10.1107/S2059798318006551
Chen, V. B. et al. MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr. D. Biol. Crystallogr. 66, 12–21 (2010).
pubmed: 20057044 doi: 10.1107/S0907444909042073
Pintilie, G. et al. Measurement of atom resolvability in cryo-EM maps with Q-scores. Nat. Methods 17, 328–334 (2020).
pubmed: 32042190 pmcid: 7446556 doi: 10.1038/s41592-020-0731-1
Shapiro, S. K. & Ehninger, D. J. Methods for the analysis and preparation of adenosylmethionine and adenosylhomocysteine. Anal. Biochem. 15, 323–333 (1966).
pubmed: 4289755 doi: 10.1016/0003-2697(66)90038-8

Auteurs

Linnea K M Blomgren (LKM)

Division of Metabolism and Children's Research Center, University Children's Hospital Zürich, University of Zürich, Zürich, CH-8032, Switzerland.

Melanie Huber (M)

Division of Metabolism and Children's Research Center, University Children's Hospital Zürich, University of Zürich, Zürich, CH-8032, Switzerland.

Sabrina R Mackinnon (SR)

Biosciences Institute, The Medical School, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.

Céline Bürer (C)

Division of Metabolism and Children's Research Center, University Children's Hospital Zürich, University of Zürich, Zürich, CH-8032, Switzerland.

Arnaud Baslé (A)

Biosciences Institute, The Medical School, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.

Wyatt W Yue (WW)

Biosciences Institute, The Medical School, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK. Wyatt.Yue@newcastle.ac.uk.
Centre for Medicines Discovery, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, OX3 7DQ, UK. Wyatt.Yue@newcastle.ac.uk.

D Sean Froese (DS)

Division of Metabolism and Children's Research Center, University Children's Hospital Zürich, University of Zürich, Zürich, CH-8032, Switzerland. Sean.Froese@kispi.uzh.ch.

Thomas J McCorvie (TJ)

Biosciences Institute, The Medical School, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK. Thomas.McCorvie@newcastle.ac.uk.

Classifications MeSH