Primary structure of the thermostable formyltetrahydrofolate synthetase from Clostridium thermoaceticum.
The complete nucleotide sequence of the Clostridium thermoaceticum formyltetrahydrofolate synthetase (FTHFS) was determined and the primary structure of the protein predicted. The gene was 1680 nucleotides long, encoding a protein of 559 amino acid residues with a calculated subunit molecular weight of 59,983. The initiation codon was UUG, with a probable ribosome binding site 11 bases upstream. A putative ATP binding domain was identified. Two Cys residues likely to be involved in subunit aggregation were tentatively identified. No characterization of the tetrahydrofolate (THF) binding domain was possible on the basis of the sequence. A high level of amino acid sequence conservation between the C. thermoaceticum FTHFS and the published sequences of C. acidiurici FTHFS and the FTHFS domains of the Saccharomyces cerevisiae C1-THF synthases was found. Of the 556 residues shared between the two clostridial sequences, 66.4% are identical. If conservative substitutions are allowed, this percentage rises to 75%. Over 47% of the residues shared between the C. thermoaceticum FTHFS and the yeast C1-THF synthases are identical, 57.4% if conservative substitutions are allowed. Hydrophobicity profiles of the C. acidiurici and C. thermoaceticum enzymes were very similar and did not support the idea that large hydrophobic domains play an important role in thermostabilizing the C. thermoaceticum FTHFS.
Department of Biological Sciences, University of South Carolina, Columbia 29208.
C R
CR
Lovell
A
A
Przybyla
L G
LG
Ljungdahl
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D003013:Clostridium / Q000201:enzymology*; D003593:Cytoplasm / Q000201:enzymology; D005574:Formate-Tetrahydrofolate Ligase / Q000032:analysis; D006358:Hot Temperature; D008025:Ligases / Q000032:analysis; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D000596:Amino Acids; D008025:Ligases; D005574:Formate-Tetrahydrofolate Ligase