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BIO DESIGN

pISSN 2288-6982
eISSN 2288-7105

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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

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.