Expression, purification, crystallization and preliminary X‐ray analysis of the PaaI‐like thioesterase SAV0944 from Staphylococcus aureus. Issue 2 (1st February 2014)

• Record Type: Journal Article
• Authors: Khandokar, Yogesh B.; Roman, Noelia; Smith, Kate M.; Srivastava, Parul; Forwood, Jade K.
• Is Part Of: Acta crystallographica. Volume 70:Issue 2(2014:Feb.)
• Page Start: 244
• Page End: 247
• Publication Date: 2014-02-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X14000338
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p> <italic>Staphylococcus aureus</italic> is the causative agent of many diseases, including meningitis, bacteraemia, pneumonia, food poisoning and toxic shock syndrome. Structural characterization of the PaaI‐like thioesterase SAV0944 (<italic>Sa</italic>PaaI) from <italic>S. aureus</italic> subsp. <italic>aureus</italic> Mu50 will aid in understanding its potential as a new therapeutic target by knowledge of its molecular details and cellular functions. Here, the recombinant expression, purification and crystallization of <italic>Sa</italic>PaaI thioesterase from <italic>S. aureus</italic> are reported. This protein initially crystallized with the ligand coenzyme A using the hanging‐drop vapour‐diffusion technique with condition No. 40 of Crystal Screen from Hampton Research at 296 K. Optimal final conditions consisting of 24% PEG 4000, 100 m<italic>M</italic> sodium citrate pH 6.5, 12% 2‐propanol gave single diffraction‐quality crystals. These crystals diffracted to beyond 2 Å resolution at the Australian Synchrotron and belonged to space group <italic>P</italic>12<sub>1</sub>1, with unit‐cell parameters <italic>a</italic> = 44.05, <italic>b</italic> = 89.05, <italic>c</italic> = 60.74 Å, β = 100.5°. Initial structure determination and refinement gave an <italic>R</italic> factor and <italic>R</italic><sub>free</sub> of 17.3 and 22.0%, respectively, confirming a positive solution in obtaining phases using molecular replacement.</p> </abstract>