Crystallization and preliminary X‐ray diffraction analysis of YhbJ from Escherichia coli, a key protein involved in the GlmYZ sRNA regulatory cascade. Issue 2 (5th February 2013)

• Record Type: Journal Article
• Authors: Resch, Marcus; Göpel, Yvonne; Görke, Boris; Ficner, Ralf
• Is Part Of: Acta crystallographica. Volume 69:Issue 2(2013:Feb.)
• Page Start: 109
• Page End: 114
• Publication Date: 2013-02-05
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309112048622
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>The protein YhbJ from <italic>Escherichia coli</italic> was previously reported to be involved in the regulation of glucosamine‐6‐phosphate synthase (GlmS) synthesis. YhbJ controls a regulatory cascade composed of the two small RNAs GlmY and GlmZ, which in turn regulate GlmS synthesis. For structural characterization, YhbJ was cloned, expressed and purified to homogeneity by <italic>Strep</italic>‐tag affinity chromatography and size‐exclusion chromatography. Multi‐angle laser light‐scattering analysis revealed its homotrimeric state in solution. The protein crystallized in two distinct trigonal crystal forms, with unit‐cell parameters <italic>a</italic> = <italic>b</italic> = 91.62, <italic>c</italic> = 352.82 Å for space group <italic>P</italic>321 and <italic>a</italic> = <italic>b</italic> = 92.72, <italic>c</italic> = 156.75 Å for one of the enantiomorphic space groups <italic>P</italic>3₁ or <italic>P</italic>3₂. Preliminary analysis of the diffraction data suggests the presence of approximately three to seven molecules per asymmetric unit. Owing to the lack of a suitable homologous model, structure determination by means of MIR and MAD methods is required.</p> </abstract>