3WUY

Crystal structure of Nit6803

Summary for 3WUY

• Entry DOI: 10.2210/pdb3wuy/pdb
• Descriptor: Nitrilase (2 entities in total)
• Functional Keywords: nitrilase, hydrolase
• Biological source: Synechocystis
• Total number of polymer chains: 2
• Total formula weight: 76613.77

Authors

Yuan, Y.A.,Yin, B.,Wang, C. (deposition date: 2014-05-09, release date: 2014-12-31, Last modification date: 2024-04-03)

Primary citation

Zhang, L.,Yin, B.,Wang, C.,Jiang, S.,Wang, H.,Yuan, Y.A.,Wei, D.
Structural insights into enzymatic activity and substrate specificity determination by a single amino acid in nitrilase from Syechocystis sp. PCC6803
J.Struct.Biol., 188:93-101, 2014

PubMed Abstract: Nitrilases are enzymes widely expressed in prokaryotes and eukaryotes that utilize a Cys–Glu–Lys catalytic triad to hydrolyze non-peptide carbon–nitrogen bonds. Nitrilase from Syechocystis sp. Strain PCC6803 (Nit6803) shows hydrolysis activity towards a broad substrate spectrum, ranging from mononitriles to dinitriles and from aromatic nitriles to aliphatic nitriles. Yet, the structural principle of the substrate specificity of this nitrilase is still unknown. We report the crystal structure of Nit6803 at 3.1 Å resolution and propose a structural mechanism of substrate selection. Our mutagenesis data exhibited that the aromaticity of the amino acid at position 146 of Nit6803 is absolutely required for its nitrilase activity towards any substrates tested. Moreover, molecular docking and dynamic simulation analysis indicated that the distance between the sulfhydryl group of the catalytic cysteine residue and the cyano carbon of the substrate plays a crucial role in determining the nitrilase catalytic activity of Nit6803 and its mutants towards different nitrile substrates.

PubMed: 25450592
DOI: 10.1016/j.jsb.2014.10.003

Experimental method

X-RAY DIFFRACTION (3.1 Å)