1XF8

Crystal Structure of Weissella viridescens FemX (Y254F) Mutant

Summary for 1XF8

• Entry DOI: 10.2210/pdb1xf8/pdb
• Related: 1LRZ 1NE9 1P4N 1XE4
• Descriptor: FemX, MAGNESIUM ION (3 entities in total)
• Functional Keywords: mutant, femx, ligase, transferase
• Biological source: Weissella viridescens
• Total number of polymer chains: 1
• Total formula weight: 38204.11

Authors

Biarrotte-Sorin, S.,Maillard, A.P.,Arthur, M.,Mayer, C. (deposition date: 2004-09-14, release date: 2005-05-31, Last modification date: 2023-10-25)

Primary citation

Maillard, A.P.,Biarrotte-Sorin, S.,Villet, R.,Mesnage, S.,Bouhss, A.,Sougakoff, W.,Mayer, C.,Arthur, M.
Structure-Based Site-Directed Mutagenesis of the UDP-MurNAc-Pentapeptide-Binding Cavity of the FemX Alanyl Transferase from Weissella viridescens
J.BACTERIOL., 187:3833-3838, 2005

PubMed Abstract: Weissella viridescens FemX (FemX(Wv)) belongs to the Fem family of nonribosomal peptidyl transferases that use aminoacyl-tRNA as the amino acid donor to synthesize the peptide cross-bridge found in the peptidoglycan of many species of pathogenic gram-positive bacteria. We have recently solved the crystal structure of FemX(Wv) in complex with the peptidoglycan precursor UDP-MurNAc-pentapeptide and report here the site-directed mutagenesis of nine residues located in the binding cavity for this substrate. Two substitutions, Lys36Met and Arg211Met, depressed FemX(Wv) transferase activity below detectable levels without affecting protein folding. Analogues of UDP-MurNAc-pentapeptide lacking the phosphate groups or the C-terminal D-alanyl residues were not substrates of the enzyme. These results indicate that Lys36 and Arg211 participate in a complex hydrogen bond network that connects the C-terminal D-Ala residues to the phosphate groups of UDP-MurNAc-pentapeptide and constrains the substrate in a conformation that is essential for transferase activity.

PubMed: 15901708
DOI: 10.1128/JB.187.11.3833-3838.2005

Experimental method

X-RAY DIFFRACTION (1.9 Å)