Site-directed mutagenesis on (serine) carboxypeptidase Y. A hydrogen bond network stabilizes the transition state by interaction with the C-terminal carboxylate group of the substrate

Biochemistry ◽  
1994 ◽  
Vol 33 (2) ◽  
pp. 508-517 ◽  
Author(s):  
Uffe H. Mortensen ◽  
S. James Remington ◽  
Klaus Breddam
2005 ◽  
Vol 187 (11) ◽  
pp. 3833-3838 ◽  
Author(s):  
Antoine P. Maillard ◽  
Sabrina Biarrotte-Sorin ◽  
Régis Villet ◽  
Stéphane Mesnage ◽  
Ahmed Bouhss ◽  
...  

ABSTRACT Weissella viridescens FemX (FemXWv) 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 FemXWv 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 FemXWv 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.


Biochemistry ◽  
2002 ◽  
Vol 41 (49) ◽  
pp. 14489-14498 ◽  
Author(s):  
Greg A. Kicska ◽  
Peter C. Tyler ◽  
Gary B. Evans ◽  
Richard H. Furneaux ◽  
Wuxian Shi ◽  
...  

1995 ◽  
Vol 117 (22) ◽  
pp. 5944-5950 ◽  
Author(s):  
Steen Bech Sorensen ◽  
Mette Raaschou-Nielsen ◽  
Uffe H. Mortensen ◽  
S. James Remington ◽  
Klaus Breddam

2021 ◽  
pp. 120431
Author(s):  
Akinori Honda ◽  
Shunta Kakihara ◽  
Shuhei Ichimura ◽  
Kazuaki Tomono ◽  
Mina Matsushita ◽  
...  

2021 ◽  
Author(s):  
Xiang-Yang Liu ◽  
Teng-Shuo Zhang ◽  
Qiu Fang ◽  
Wei-Hai Fang ◽  
Leticia González ◽  
...  

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