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Abstract

Most organisms possess bifunctional FolD [5,10-methylenetetrahydrofolate (5,10-CH-THF) dehydrogenase-cyclohydrolase] to generate NADPH and 10-formyltetrahdrofolate (10-CHO-THF) required in various metabolic steps. In addition, some organisms including possess another protein, Fhs (formyltetrahydrofolate synthetase), to synthesize 10-CHO-THF. Here, we show that unlike the bifunctional FolD of (FolD), and contrary to its annotated bifunctional nature, FolD (FolD) is a monofunctional 5,10-CH-THF dehydrogenase. The dehydrogenase activity of FolD is about five times more efficient than that of FolD. The 5,10-methenyltetrahydrofolate (5,10-CH-THF) cyclohydrolase activity in is provided by another protein, FchA (5,10-CH-THF cyclohydrolase), whose cyclohydrolase activity is ∼10 times more efficient than that of FolD. Kinetic parameters for Fhs were also determined for utilization of all of its substrates. Both FolD and FchA are required to substitute for the single bifunctional FolD in . The simultaneous presence of FolD and FchA is also necessary to rescue an deletion strain (harbouring Fhs support) for its formate and glycine auxotrophies, and to alleviate its susceptibility to trimethoprim (an antifolate drug) or UV light. The presence of the three clostridial proteins (FolD, FchA and Fhs) is required to maintain folate homeostasis in the cell.

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2016-01-01
2021-07-26
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