Expression, crystallization and preliminary crystallographic analysis of SufE (XAC2355) from Xanthomonas axonopodis pv. citri.
Xanthomonas axonopodis pv. citri (Xac) SufE (XAC2355) is a member of a family of bacterial proteins that are conserved in several pathogens and phytopathogens. The Escherichia coli suf operon is involved in iron-sulfur cluster biosynthesis under iron-limitation and stress conditions. It has recently been demonstrated that SufE and SufS form a novel two-component cysteine desulfarase in which SufS catalyses the conversion of L-cysteine to L-alanine, forming a protein-bound persulfide intermediate. The S atom is then transferred to SufE, from which it is subsequently transferred to target molecules or reduced to sulfide in solution. Here, the cloning, expression, crystallization and phase determination of Xac SufE crystals are described. Recombinant SufE was crystallized in space group P2(1)2(1)2(1) and diffracted to 1.9 A resolution at a synchrotron source. The unit-cell parameters are a = 45.837, b = 58.507, c = 98.951 A, alpha = beta = gamma = 90 degrees. The calculated Matthews coefficient indicated the presence of two molecules in the asymmetric unit. Phasing was performed by molecular-replacement using E. coli SufE as a model (PDB code 1mzg) and an interpretable map was obtained.
Guzzo
Cristiane R
CR
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, CEP 05508-000, São Paulo, SP, Brazil.
Silva
Lucicleide R
LR
Galvão-Botton
Leonor M P
LM
Barbosa
João A R G
JA
Farah
Chuck S
CS
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D013437:Carbon-Sulfur Lyases / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D014974:Xanthomonas / Q000737:chemistry*
D001426:Bacterial Proteins; D013437:Carbon-Sulfur Lyases; C080546:cysteine desulfurase