Putative thioredoxin Trx1 from Thermosipho africanus strain TCF52B: expression, purification and structural determination using S‐SAD. Issue 6 (1st June 2016)

• Record Type: Journal Article
• Authors: Sahtout, Naheda; Kuttiyatveetil, Jijin R. A.; Fodje, Michel; Sanders, David A. R.
• Is Part Of: Acta crystallographica. Volume 72:Issue 6(2016:Jun.)
• Page Start: 443
• Page End: 447
• Publication Date: 2016-06-01
• Subjects: thioredoxin -- Thermosipho africanus -- thermophile -- sulfur -- SAD; Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X16007214
• Languages: English
• ISSNs: 2053-230X

Abstract

Abstract : Crystallographic analysis of the putative thioredoxin Trx1 from T. africanus strain TCF52B, which has low sequence identity to its closest homologues, was completed using sulfur single‐wavelength anomalous dispersion analysis. Abstract : Thioredoxin is a small ubiquitous protein that plays a role in many biological processes. A putative thioredoxin, Trx1, from Thermosipho africanus strain TCF52B, which has low sequence identity to its closest homologues, was successfully cloned, overexpressed and purified. The protein was crystallized using the microbatch‐under‐oil technique at 289 K in a variety of conditions; crystals grown in 0.2 M MgCl2, 0.1 M bis‐tris pH 6.5, 25%( w / v ) PEG 3350, which grew as irregular trapezoids to maximum dimensions of 1.2 × 1.5 × 0.80 mm, were used for sulfur single‐wavelength anomalous dispersion analysis. The anomalous sulfur signal could be detected to 2.83 Å resolution using synchrotron radiation on the 08B1‐1 beamline at the Canadian Light Source. The crystals belonged to space group P 21 21 21, with unit‐cell parameters a = 40.6, b = 41.5, c = 56.4 Å, α = β = γ = 90.0°.