(Beta alpha)8-barrel proteins of tryptophan biosynthesis in the hyperthermophile Thermotoga maritima.
To better understand the evolution of a key metabolic pathway, we have sequenced the trpCFBA gene cluster of the hyperthermophilic bacterium Thermotoga maritima. The genes were cloned by complementation in vivo of trp deletion strains of Escherichia coli. The new sequences, together with earlier findings, establish that the trp operon of T.maritima has the order trpE(G.D)CFBA, which might represent the ancestral organization of the tryptophan operon. Heterologous expression of the trp(G.D) and trpC genes in E.coli and N-terminal sequencing of their polypeptide products showed that their translation is initiated at the rate start codons TTG and ATC, respectively. Consequently, the N-terminus of the trp(G.D) fusion protein is 43 residues shorter than previously postulated. Amino acid composition and sequence analyses of the protein products of T.maritima trpC (indoleglycerol phosphate synthase), trpF (phosphoribosyl anthranilate isomerase) and trpA (alpha-subunit of tryptophan synthase) suggest that these thermostable (beta alpha)8-barrel proteins may be stabilized by additional salt bridges, compared with the mesostable forms. Another notable feature is the predicted lack of the N-terminal helix alpha 0 in the alpha-subunit of tryptophan synthase.
Sterner
R
R
Abteilung für Biophysikalische Chemie, Universität Basel, Switzerland.
Dahm
A
A
Darimont
B
B
Ivens
A
A
Liebl
W
W
Kirschner
K
K
D019747:Aldose-Ketose Isomerases; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002238:Carbohydrate Epimerases / Q000235:genetics*; D003001:Cloning, Molecular; D018387:Codon, Initiator; D004795:Enzyme Stability / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology / Q000235:genetics*; D006358:Hot Temperature; D007209:Indole-3-Glycerol-Phosphate Synthase / Q000235:genetics*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics*; D009876:Operon / Q000235:genetics; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA; D014364:Tryptophan / Q000096:biosynthesis*; D014367:Tryptophan Synthase / Q000235:genetics*
D001426:Bacterial Proteins; D018387:Codon, Initiator; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; C052909:phosphoribosylanthranilate isomerase-indoleglycerol-phosphate synthase; D014364:Tryptophan; D007209:Indole-3-Glycerol-Phosphate Synthase; D014367:Tryptophan Synthase; D002238:Carbohydrate Epimerases; D019747:Aldose-Ketose Isomerases