10-Formyltetrahydrofolate synthetase. Evidence for a conformational change in the enzyme upon binding of tetrahydropteroylpolyglutamates.
The 10-formyltetrahydrofolate synthetase domain of the trifunctional enzyme C1-tetrahydrofolate synthase appears to undergo a conformational change in the presence of tetrahydropteroylpolyglutamates, MgATP, and ammonium ion. The binding of these ligands increases the denaturation temperature of the enzyme by 12 degrees C, abolishes the cold lability of the enzyme, and alters its susceptibility to digestion by chymotrypsin. The results suggest that a conformational change is dependent upon binding of the third glutamate residue of tetrahydropteroylpolyglutamates and the beta-phosphoryl group of MgATP. The Km values for MgATP and formate are lowered 3.6- and 520-fold, respectively, when tetrahydropteroyltriglutamate is used as the substrate in place of tetrahydropteroylmonoglutamate. A sensitive coupled assay involving C1-tetrahydrofolate synthase and serine hydroxymethyltransferase was developed to determine the activity of 10-formyltetrahydrofolate synthetase. The assay gives linear rates with the tetrahydropteroylpolyglutamates as substrates but not with the monoglutamate form.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000619:Aminohydrolases / Q000378:metabolism*; D000641:Ammonia / Q000378:metabolism; D000818:Animals; D002152:Calorimetry, Differential Scanning; D005574:Formate-Tetrahydrofolate Ligase / Q000378:metabolism*; D012696:Glycine Hydroxymethyltransferase; D007700:Kinetics; D008025:Ligases / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D045064:Methenyltetrahydrofolate Cyclohydrolase; D008754:Methylenetetrahydrofolate Dehydrogenase (NADP) / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D011817:Rabbits; D013696:Temperature; D013763:Tetrahydrofolates / Q000378:metabolism*
D009097:Multienzyme Complexes; D013763:Tetrahydrofolates; C030371:5,6,7,8-tetrahydrofolic acid; C021836:formyl-methenyl-methylenetetrahydrofolate synthetase; D000641:Ammonia; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; D008754:Methylenetetrahydrofolate Dehydrogenase (NADP); D012696:Glycine Hydroxymethyltransferase; D000619:Aminohydrolases; D045064:Methenyltetrahydrofolate Cyclohydrolase; D008025:Ligases; D005574:Formate-Tetrahydrofolate Ligase; D008274:Magnesium