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Expression, purification, crystallization and preliminary X‐ray diffraction analysis of the C‐terminal NHL domain of human TRIM2
Author(s) -
Guan Xiaotao,
Li Jun,
Lü Xingru,
Dong Yu,
Chen Weimin,
Li Xuemei
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14008127
Subject(s) - crystallization , terminal (telecommunication) , crystallography , domain (mathematical analysis) , c terminus , materials science , diffraction , x ray crystallography , x ray , chemistry , optics , computer science , chemical engineering , physics , engineering , biochemistry , mathematics , telecommunications , mathematical analysis , amino acid
The tripartite motif‐containing protein 2 (TRIM2) functions as an E3 ubiquitin ligase. Loss of function of TRIM2 has been shown to result in early‐onset axonal neuropathy. As a member of the TRIM–NHL family of proteins, TRIM2 has a conserved modular architecture that includes N‐terminal RING finger and B‐box domains, a middle coiled‐coil domain and a C‐terminal NHL domain. To characterize the functional role of its NHL domain from the perspective of structural biology, a truncation of human TRIM2 (residues 465–744) was expressed, purified and crystallized. Rod‐shaped crystals were obtained that diffracted X‐rays to 1.7 Å resolution. The crystals belonged to space group P 2 1 , with unit‐cell parameters a = 43.6, b = 76.4, c = 107.4 Å, α = 90.0, β = 94.0, γ = 90.0°. A Matthews coefficient of 1.97 Å 3 Da −1 , corresponding to a solvent content of 37.6%, indicated the presence of three molecules per asymmetric unit, which was further confirmed by the phasing solution from molecular replacement.