Purification, crystallization and preliminary X‐ray analysis of the periplasmic haem‐binding protein HutB from Vibrio cholerae. Issue 4 (1st April 2015)

• Record Type: Journal Article
• Authors: Agarwal, Shubhangi; Biswas, Maitree; Dasgupta, Jhimli
• Is Part Of: Acta crystallographica. Volume 71:Issue 4(2015:Apr.)
• Page Start: 401
• Page End: 404
• Publication Date: 2015-04-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X15003660
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>The mechanism of haem transport across the inner membrane of pathogenic bacteria is currently insufficiently understood at the molecular level and no information is available for this process in <italic>Vibrio cholerae</italic>. To obtain structural insights into the periplasmic haem‐binding protein HutB from <italic>V. cholerae</italic> (VcHutB), which is involved in haem transport through the HutBCD haem‐transport system, at the atomic level, VcHutB was cloned, overexpressed and crystallized using 1.6 <italic>M</italic> ammonium sulfate as a precipitant at pH 7.0. X‐ray diffraction data were collected to 2.4 Å resolution on the RRCAT PX‐BL‐21 beamline at the Indus‐2 synchrotron, Indore, India. The crystals belonged to space group <italic>P</italic>4₃2₁2, with unit‐cell parameters <italic>a</italic> = <italic>b</italic> = 62.88, <italic>c</italic> = 135.8 Å. Matthews coefficient calculations indicated the presence of one monomer in the asymmetric unit, with an approximate solvent content of 45.02%. Molecular‐replacement calculations with <italic>Phaser</italic> confirmed the presence of a monomer in the asymmetric unit.</p> </abstract>