title: Proton nuclear magnetic resonance studies on the wild-type and single amino acid substituted tryptophan synthase alpha-subunits.
abstract: In order to elucidate the effect of single amino acid substitutions on the conformation of the tryptophan synthase alpha-subunit from Escherichia coli in solution, 1H NMR spectra of the wild-type and mutant proteins were measured at various pHs. Two of the four His C2-proton resonances of the alpha-subunit were assigned to two His residues at positions 92 and 146 by using a mutant protein with Thr substituted for the His at position 92. The replacement did not affect the conformation of the protein significantly. The proton resonances of all the Tyr residues in the aromatic region could be picked up from other resonance peaks, employing the wild-type alpha-subunit deuterated at all of the Phe residues. On comparison of the spectra of the wild-type protein with those of the mutant protein with Met substituted for the Glu at position 49, it was concluded that the substitution affects only the residues close to the substituted residue at acidic pH but that a larger part of the protein is affected at alkaline pH. NOE experiments showed that the five Tyr residues, four of which are located in the proximity of position 49, are close to one another. The present results are discussed in the light of the conformational stability of the protein.
authors.affiliation: Institute for Protein Research, Osaka University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yutani
authors.forename: H
authors.initials: H
authors.lastname: Akutsu
authors.forename: K
authors.initials: K
authors.lastname: Ogasahara
authors.forename: T
authors.initials: T
authors.lastname: Tsujita
authors.forename: Y
authors.initials: Y
authors.lastname: Kyogoku
mesh_terms: D000596:Amino Acids; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008715:Methionine; D009154:Mutation; D011487:Protein Conformation; D014367:Tryptophan Synthase / Q000235:genetics*
chemical_list: D000596:Amino Acids; D046911:Macromolecular Substances; D008715:Methionine; D014367:Tryptophan Synthase