Characterization, crystallization and preliminary X‐ray crystallographic analysis of the Uba5 fragment necessary for high‐efficiency activation of Ufm1. Issue 6 (1st June 2014)

• Record Type: Journal Article
• Authors: Xie, Shutao
• Is Part Of: Acta crystallographica. Volume 70:Issue 6(2014:Jun.)
• Page Start: 765
• Page End: 768
• Publication Date: 2014-06-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X14008826
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Uba5 is the smallest ubiquitin‐like molecule‐activating enzyme and contains an adenylation domain and a C‐terminal region. This enzyme only exists in multicellular organisms. The mechanism through which the enzyme recognizes and activates ubiquitin‐fold modifier 1 (Ufm1) remains unknown. In this study, Uba5 adenylation domains with different C‐terminal region lengths were cloned, expressed and purified. The results of an <italic>in vitro</italic> truncation assay suggest that Uba5 residues 57–363 comprise the minimal fragment required for the high‐efficiency activation of Ufm1. Crystallization of Uba5 residues 57–363 was performed at 277 K using PEG 3350 as the precipitant, and crystals optimized by microseeding diffracted to 2.95 Å resolution, with unit‐cell parameters <italic>a</italic> = <italic>b</italic> = 97.66, <italic>c</italic> = 144.83 Å, α = β = 90, γ = 120°. There is one molecule in the asymmetric unit; the Matthews coefficient and the solvent content were calculated to be 2.93 ų Da⁻¹ and 58.1%, respectively.</p> </abstract>