Abstract
Elevated plasma homocysteine levels are associated with increased risk for cardiovascular disease and neural tube defects in humans. Folate treatment decreases homocysteine levels and dramatically reduces the incidence of neural tube defects. The flavoprotein methylenetetrahydrofolate reductase (MTHFR) is a likely target for these actions of folate. The most common genetic cause of mildly elevated plasma homocysteine in humans is the MTHFR polymorphism A222V (base change C677→T). The X-ray analysis of E. coli MTHFR, reported here, provides a model for the catalytic domain that is shared by all MTHFRs. This domain is a β8α8 barrel that binds FAD in a novel fashion. Ala 177, corresponding to Ala 222 in human MTHFR, is near the bottom of the barrel and distant from the FAD. The mutation A177V does not affect Km or kcat but instead increases the propensity for bacterial MTHFR to lose its essential flavin cofactor. Folate derivatives protect wild-type and mutant E. coli enzymes against flavin loss, and protect human MTHFR and the A222V mutant against thermal inactivation, suggesting a mechanism by which folate treatment reduces homocysteine levels.
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Acknowledgements
We thank J. Shih (Lilly Corporation) for the gift of the pentaglutamate form of 5,10-dideazatetrahydrofolate. We are indebted to C. Johnson (Calorimetry Sciences), for performing the differential-scanning calorimetry experiments shown in Fig. 4. We also thank A. Gafni for allowing us to use his fluorimeter and D. Ballou and V. Massey for assistance with the stopped-flow experiments. This work was supported by National Institute of General Medical Sciences grants to R.M and M.L., by NIH postdoctoral (B.G.) and predoctoral (C.S.) fellowships, by an NIH Cellular & Molecular Biology Training Grant (C.S.), and the Medical Research Council of Canada (R.R.) and by the McGill University Mary Louise Taylor predoctoral fellowship (P.T.).
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Guenther, B., Sheppard, C., Tran, P. et al. The structure and properties of methylenetetrahydrofolate reductase from Escherichia coli suggest how folate ameliorates human hyperhomocysteinemia . Nat Struct Mol Biol 6, 359–365 (1999). https://doi.org/10.1038/7594
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DOI: https://doi.org/10.1038/7594
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