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Expression, purification, crystallization and preliminary X‐ray analysis of the PaaI‐like thioesterase SAV0944 from Staphylococcus aureus
Author(s) -
Khandokar Yogesh B.,
Roman Noelia,
Smith Kate M.,
Srivastava Parul,
Forwood Jade K.
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/s2053230x14000338
Subject(s) - staphylococcus aureus , crystallization , chemistry , molecular replacement , toxin , exotoxin , crystallography , microbiology and biotechnology , biology , biochemistry , bacteria , genetics , organic chemistry
Staphylococcus aureus 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 ( Sa PaaI) from S. aureus subsp. aureus 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 Sa PaaI thioesterase from S. aureus 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 M 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 P 12 1 1, with unit‐cell parameters a = 44.05, b = 89.05, c = 60.74 Å, β = 100.5°. Initial structure determination and refinement gave an R factor and R free of 17.3 and 22.0%, respectively, confirming a positive solution in obtaining phases using molecular replacement.