title: The fMet-tRNA binding domain of translational initiation factor IF2: role and environment of its two Cys residues.
abstract: Mutations of the cysteines (positions 668 and 714) were generated in the IF2 C domain of Bacillus stearothermophilus translation initiation factor IF2. The corresponding proteins were characterized functionally and structurally. Most (yet not all) amino acid replacements at both positions resulted in severe reduction of the fMet-tRNA binding activity of IF2 C without grossly altering its structure. Our work demonstrates that: (a) both Cys residues are buried within an hydrophobic core and not accessible to protonation or chemical substitution, (b) neither Cys is functionally essential and (c) both Cys residues are located near the active site, probably without participating directly in fMet-tRNA binding.
authors.affiliation: Max-Delbrück-Centre for Molecular Medicine, D-13092, Berlin, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Misselwitz
authors.forename: K
authors.initials: K
authors.lastname: Welfle
authors.forename: C
authors.initials: C
authors.lastname: Krafft
authors.forename: H
authors.initials: H
authors.lastname: Welfle
authors.forename: L
authors.initials: L
authors.lastname: Brandi
authors.forename: E
authors.initials: E
authors.lastname: Caserta
authors.forename: C O
authors.initials: CO
authors.lastname: Gualerzi
mesh_terms: D001665:Binding Sites; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D001411:Geobacillus stearothermophilus / Q000235:genetics / Q000378:metabolism*; D019791:Guanidine / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010448:Peptide Initiation Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D039881:Prokaryotic Initiation Factor-2; D011487:Protein Conformation; D011489:Protein Denaturation; D012358:RNA, Transfer, Met / Q000378:metabolism*; D013059:Spectrum Analysis, Raman
chemical_list: D010448:Peptide Initiation Factors; D039881:Prokaryotic Initiation Factor-2; D012358:RNA, Transfer, Met; C024988:tRNA, formylmethionine-; D019791:Guanidine; D003545:Cysteine