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BIO DESIGN

pISSN 2288-6982
eISSN 2288-7105

Article

Article

Crystallization

Bio Design 2020; 8(1): 20-23

Published online March 30, 2020

https://doi.org/10.34184/kssb.2020.8.1.20

© Korean Society for Structural Biology

Purification, crystallization, and X-ray crystallographic analysis of Nit2 from Kluyveromyces lactis

Chaewon Jin1, Hyeonseok Jin1, Bradley Quade2,* and Jeong Ho Chang1,2,*

1Department of Biology Education, Kyungpook National University, Daegu 41566, Republic of Korea
2Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA

Correspondence to: *jhcbio@knu.ac.kr, bradley.quade@utsouthwestern.edu

Received: November 5, 2019; Revised: March 15, 2020; Accepted: March 15, 2020

Abstract

The nitrilase-like (NIT) protein subfamily is a member of the nitrilase superfamily, which exhibits CN hydrolase activity. NIT proteins are highly conserved, and mammals have Nit1 and Nit2. Nit proteins are putative tumor suppressors; however, the enzymatic activities of Nit1 and Nit2 are different. Nit1 is a metabolite repair enzyme, whereas Nit2 is an ω-amidase. In this study, in order to understand the structural features and functional diversity of Nit proteins, Nit2 from Kluyveromyces lactis was crystallized, and X-ray diffraction data were obtained at a resolution of 2.2 Å. The crystal of K. lactis Nit2 (KlNit2) belonged to the space group C2, with the unit cell parameters a = 79.0 Å, b = 211.1 Å, c = 89.4 Å, α = γ = 90°, β = 112.1°. The crystal contained four macromolecules in the asymmetric unit.