Crystallization and preliminary X-ray analysis of a truncated mutant of yeast nuclear thiol peroxidase, a novel atypical 2-Cys peroxiredoxin.
Saccharomyces cerevisiae nTPx is a thioredoxin-dependent thiol peroxidase that is localized in the nucleus. nTPx belongs to the C-type atypical 2-Cys peroxiredoxin family members, which are frequently called BCPs or PrxQs. A double mutant (C107S/C112S) of nTPx overexpressed in Escherichia coli was spontaneously degraded upon freezing and thawing and its truncated form (residues 57-215; MW = 17837 Da) was crystallized with PEG 3350 and mercury(II) acetate as precipitants using the hanging-drop vapour-diffusion method. Diffraction data were collected to 1.8 A resolution using X-ray synchrotron radiation. The crystals belong to the trigonal space group P3(2), with unit-cell parameters a = b = 37.54, c = 83.26 A. The asymmetric unit contains one molecule of truncated mutant nTPx, with a corresponding VM of 1.91 A3 Da(-1) and a solvent content of 35.5%.
Choi
Jongkeun
J
Department of Chemistry, Seoul National University, Seoul 151-742, South Korea.
Choi
Soonwoong
S
Choi
Jungwon
J
Cha
Mee-Kyung
MK
Kim
Il-Han
IH
Shin
Whanchul
W
D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry*; D004058:Diffusion; D004926:Escherichia coli / Q000378:metabolism; D008628:Mercury / Q000737:chemistry; D009154:Mutation; D010544:Peroxidases / Q000737:chemistry*; D054464:Peroxiredoxins; D011092:Polyethylene Glycols / Q000737:chemistry; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D017356:Synchrotrons; D013879:Thioredoxins / Q000737:chemistry; D014961:X-Ray Diffraction; D014965:X-Rays
D029701:Saccharomyces cerevisiae Proteins; D012997:Solvents; D011092:Polyethylene Glycols; D013879:Thioredoxins; D010544:Peroxidases; C496253:Tsa1 protein, S cerevisiae; D054464:Peroxiredoxins; D008628:Mercury; D003545:Cysteine; C029823:mercuric acetate