Dihydrofolate reductase
From Proteopedia
The enzyme dihydrofolate reductase (DHFR) occurs in all organisms and has been particularly well-studied in the bacterium Escherichia coli and in humans[1][2][3]. It catalyzes the reduction of dihydrofolate to tetrahydrofolate, with NADPH acting as hydride donor. The human enzyme is a target for developing inhibitors used in anti-cancer chemotherapies[4], while the bacterial enzymes are targets for developing inhibitors as antibiotics. DHFR is a model enzyme for studying the kinetics, mechanism, and inhibition of enzymatic reactions and the underlying structure and conformational dynamics.
Contents |
DHFR occurs in all organisms and most cells
DHFR is found in all organisms. Some bacteria acquire resistance to DHFR inhibitors through expressing a second form of DHFR coded on a plasmid. The enzymes from E. coli (ecDHFR) and humans (hDHFR) have similar folds, while the plasmid-encoded enzyme has an unrelated fold. In humans, DHFR is expressed in most tissues[1], and there are two genes, DHFR and DHFR2/DHFRL1, the latter targeted to mitochondria[5]. Mice and rats lack the second gene but also show DHFR activity in mitochondria[6].
Reactions catalyzed
The reaction catalyzed by DHFR reduces a double bond in dihydrofolate (DHF) to form tetrahydrofolate (THF) by transfering a hydride from nicotinamide adenine dinucleotide phosphate (NADPH)
Dihydrofolate reductase (DHFR, 1.5.1.3 [2]) is an enzyme which uses the co-factor NADPH as electron donor. It catalyzes the reduction of dihydrofolic acid (DHF) to tetrahydrofolic acid (THF) as NADPH is oxidized to NADP+. The mammalian enzymes also accept folic acid as a substrate, reducing it to THF. This allows the use of folic acid, which is easier to synthesize than DHF or THF, to fortify food.[7][8]. Some bacterial enzymes also accept folic acid as a substrate [9] but it acts as a competitive inhibitor in the E. coli enzyme.
The folate is a form of the essential vitamin B9. Folate is not part of our natural diet (it contains dihydrofolate and tetrahydrofolate, sometimes as a poly-glutamate conjugate) but is bioavailable and simpler to synthesize.
Relevance
Tetrahydrofolate (THF) is an essential cofactor of one-carbon metabolism[10][11].Structure and Function
See also
- An overview of pathways involving folate and S-adenosyl methioine: One-carbon metabolism
- Another page on DHFR: Molecular Playground/DHFR
- Fighting malaria with antifolates" Malarial Dihydrofolate Reductase as Drug Target.
- Overview of enzymes targeted in cancer therapy: Cancer.
3D Structures of Dihydrofolate reductase
Dihydrofolate reductase 3D structures
Acknowledgements
This page was revised as part of the Spring 2022 Biochemistry II course at WSU. Thanks go to Kia, Anna, Shaylie and Michael for helpful suggestions to improve the page.
References
- ↑ Schnell JR, Dyson HJ, Wright PE. Structure, dynamics, and catalytic function of dihydrofolate reductase. Annu Rev Biophys Biomol Struct. 2004;33:119-40. doi:, 10.1146/annurev.biophys.33.110502.133613. PMID:15139807 doi:http://dx.doi.org/10.1146/annurev.biophys.33.110502.133613
- ↑ https://en.wikipedia.org/wiki/Dihydrofolate_reductase
- ↑ https://pdb101.rcsb.org/motm/34
- ↑ Raimondi MV, Randazzo O, La Franca M, Barone G, Vignoni E, Rossi D, Collina S. DHFR Inhibitors: Reading the Past for Discovering Novel Anticancer Agents. Molecules. 2019 Mar 22;24(6). pii: molecules24061140. doi:, 10.3390/molecules24061140. PMID:30909399 doi:http://dx.doi.org/10.3390/molecules24061140
- ↑ McEntee G, Minguzzi S, O'Brien K, Ben Larbi N, Loscher C, O'Fagain C, Parle-McDermott A. The former annotated human pseudogene dihydrofolate reductase-like 1 (DHFRL1) is expressed and functional. Proc Natl Acad Sci U S A. 2011 Sep 13;108(37):15157-62. doi:, 10.1073/pnas.1103605108. Epub 2011 Aug 26. PMID:21876184 doi:http://dx.doi.org/10.1073/pnas.1103605108
- ↑ Hughes L, Carton R, Minguzzi S, McEntee G, Deinum EE, O'Connell MJ, Parle-McDermott A. An active second dihydrofolate reductase enzyme is not a feature of rat and mouse, but they do have activity in their mitochondria. FEBS Lett. 2015 Jul 8;589(15):1855-62. doi: 10.1016/j.febslet.2015.05.017. Epub, 2015 May 14. PMID:25980602 doi:http://dx.doi.org/10.1016/j.febslet.2015.05.017
- ↑ Choi JH, Yates Z, Veysey M, Heo YR, Lucock M. Contemporary issues surrounding folic Acid fortification initiatives. Prev Nutr Food Sci. 2014 Dec;19(4):247-60. doi: 10.3746/pnf.2014.19.4.247. Epub, 2014 Dec 31. PMID:25580388 doi:http://dx.doi.org/10.3746/pnf.2014.19.4.247
- ↑ https://ods.od.nih.gov/factsheets/Folate-HealthProfessional/
- ↑ Loveridge EJ, Hroch L, Hughes RL, Williams T, Davies RL, Angelastro A, Luk LY, Maglia G, Allemann RK. Reduction of Folate by Dihydrofolate Reductase from Thermotoga maritima. Biochemistry. 2017 Apr 4;56(13):1879-1886. doi: 10.1021/acs.biochem.6b01268. Epub, 2017 Mar 24. PMID:28319664 doi:http://dx.doi.org/10.1021/acs.biochem.6b01268
- ↑ Fox JT, Stover PJ. Folate-mediated one-carbon metabolism. Vitam Horm. 2008;79:1-44. doi: 10.1016/S0083-6729(08)00401-9. PMID:18804690 doi:http://dx.doi.org/10.1016/S0083-6729(08)00401-9
- ↑ doi: https://dx.doi.org/10.1007/978-1-4020-2400-9_12
- ↑ Ivanetich KM, Santi DV. Thymidylate synthase-dihydrofolate reductase in protozoa. Exp Parasitol. 1990 Apr;70(3):367-71. PMID:2178951
- ↑ Liu CT, Francis K, Layfield JP, Huang X, Hammes-Schiffer S, Kohen A, Benkovic SJ. Escherichia coli dihydrofolate reductase catalyzed proton and hydride transfers: temporal order and the roles of Asp27 and Tyr100. Proc Natl Acad Sci U S A. 2014 Dec 23;111(51):18231-6. doi:, 10.1073/pnas.1415940111. Epub 2014 Dec 1. PMID:25453098 doi:http://dx.doi.org/10.1073/pnas.1415940111
- ↑ Sawaya MR, Kraut J. Loop and subdomain movements in the mechanism of Escherichia coli dihydrofolate reductase: crystallographic evidence. Biochemistry. 1997 Jan 21;36(3):586-603. PMID:9012674 doi:http://dx.doi.org/10.1021/bi962337c
- ↑ Wan Q, Bennett BC, Wilson MA, Kovalevsky A, Langan P, Howell EE, Dealwis C. Toward resolving the catalytic mechanism of dihydrofolate reductase using neutron and ultrahigh-resolution X-ray crystallography. Proc Natl Acad Sci U S A. 2014 Dec 1. pii: 201415856. PMID:25453083 doi:http://dx.doi.org/10.1073/pnas.1415856111
- ↑ Stojkovic V, Perissinotti LL, Willmer D, Benkovic SJ, Kohen A. Effects of the donor-acceptor distance and dynamics on hydride tunneling in the dihydrofolate reductase catalyzed reaction. J Am Chem Soc. 2012 Jan 25;134(3):1738-45. doi: 10.1021/ja209425w. Epub 2012 Jan , 17. PMID:22171795 doi:http://dx.doi.org/10.1021/ja209425w
- ↑ doi: https://dx.doi.org/10.3390/quantum3010006
- ↑ Wrobel A, Arciszewska K, Maliszewski D, Drozdowska D. Trimethoprim and other nonclassical antifolates an excellent template for searching modifications of dihydrofolate reductase enzyme inhibitors. J Antibiot (Tokyo). 2020 Jan;73(1):5-27. doi: 10.1038/s41429-019-0240-6. Epub, 2019 Oct 2. PMID:31578455 doi:http://dx.doi.org/10.1038/s41429-019-0240-6
- ↑ Estrada A, Wright DL, Anderson AC. Antibacterial Antifolates: From Development through Resistance to the Next Generation. Cold Spring Harb Perspect Med. 2016 Aug 1;6(8). pii: cshperspect.a028324. doi:, 10.1101/cshperspect.a028324. PMID:27352799 doi:http://dx.doi.org/10.1101/cshperspect.a028324
- ↑ Capasso C, Supuran CT. Sulfa and trimethoprim-like drugs - antimetabolites acting as carbonic anhydrase, dihydropteroate synthase and dihydrofolate reductase inhibitors. J Enzyme Inhib Med Chem. 2014 Jun;29(3):379-87. doi:, 10.3109/14756366.2013.787422. Epub 2013 Apr 29. PMID:23627736 doi:http://dx.doi.org/10.3109/14756366.2013.787422
- ↑ Narayana N, Matthews DA, Howell EE, Nguyen-huu X. A plasmid-encoded dihydrofolate reductase from trimethoprim-resistant bacteria has a novel D2-symmetric active site. Nat Struct Biol. 1995 Nov;2(11):1018-25. PMID:7583655
Proteopedia Page Contributors and Editors (what is this?)
Michal Harel, Karsten Theis, Alexander Berchansky, Joel L. Sussman, Tzvia Selzer, Jaime Prilusky, Eric Martz, Eran Hodis, David Canner