title: Prediction of the tertiary structure of the alpha-subunit of tryptophan synthase.
abstract: The tertiary structure of the alpha-subunit of tryptophan synthase was proposed using a combination of experimental data and computational methods. The vacuum-ultraviolet circular dichroism spectrum was used to assign the protein to the alpha/beta-class of supersecondary structures. The two-domain structure of the alpha-subunit (Miles et al.: Biochemistry 21:2586, 1982; Beasty and Matthews: Biochemistry 24:3547, 1985) eliminated consideration of a barrel structure and focused attention on a beta-sheet structure. An algorithm (Cohen et al.: Biochemistry 22:4894, 1983) was used to generate a secondary structure prediction that was consistent with the sequence data of the alpha-subunit from five species. Three potential secondary structures were then packed into tertiary structures using other algorithms. The assumption of nearest neighbors from second-site revertant data eliminated 97% of the possible tertiary structures; consideration of conserved hydrophobic packing regions on the beta-sheet eliminated all but one structure. The native structure is predicted to have a parallel beta-sheet flanked on both sides by alpha-helices, and is consistent with the available data on chemical cross-linking, chemical modification, and limited proteolysis. In addition, an active site region containing appropriate residues could be identified as well as an interface for beta 2-subunit association. The ability of experimental data to facilitate the prediction of protein structure is discussed.
authors.affiliation: Department of Chemistry, Pennsylvania State University, University Park 16802.
authors.forename: M R
authors.initials: MR
authors.lastname: Hurle
authors.forename: C R
authors.initials: CR
authors.lastname: Matthews
authors.forename: F E
authors.initials: FE
authors.lastname: Cohen
authors.forename: I D
authors.initials: ID
authors.lastname: Kuntz
authors.forename: A
authors.initials: A
authors.lastname: Toumadje
authors.forename: W C
authors.initials: WC
authors.lastname: Johnson
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002942:Circular Dichroism; D003545:Cysteine; D004926:Escherichia coli / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D014367:Tryptophan Synthase
chemical_list: D014367:Tryptophan Synthase; D003545:Cysteine