Crystallization and preliminary crystallographic characterization of LmACR2, an arsenate/antimonate reductase from Leishmania major.
Arsenic is present in the biosphere owing either to the presence of pesticides and herbicides used in agricultural and industrial activities or to leaching from geological formations. The health effects of prolonged exposure to arsenic can be devastating and may lead to various forms of cancer. Antimony(V), which is chemically very similar to arsenic, is used instead in the treatment of leishmaniasis, an infection caused by the protozoan parasite Leishmania sp.; the reduction of pentavalent antimony contained in the drug Pentostam to the active trivalent form arises from the presence in the Leishmania genome of a gene, LmACR2, coding for the protein LmACR2 (14.5 kDa, 127 amino acids) that displays weak but significant sequence similarity to the catalytic domain of Cdc25 phosphatase and to rhodanese enzymes. For structural characterization, LmACR2 was overexpressed, purified to homogeneity and crystallized in a trigonal space group (P321 or P3(1)21/P3(2)21). The protein crystallized in two distinct trigonal crystal forms, with unit-cell parameters a = b = 111.0, c = 86.1 A and a = b = 111.0, c = 175.6 A, respectively. At a synchrotron beamline, the diffraction pattern extended to a resolution limit of 1.99 A.
Bisacchi
Davide
D
Bioinformatics and Structural Proteomics, IST-National Cancer Research Institute, Genova, Italy.
Zhou
Yao
Y
Rosen
Barry P
BP
Mukhopadhyay
Rita
R
Bordo
Domenico
D
D000595:Amino Acid Sequence; D000818:Animals; D000965:Antimony / Q000737:chemistry / Q000378:metabolism; D053501:Arsenite Transporting ATPases; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D016623:Ion Pumps / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D018320:Leishmania major / Q000737:chemistry / Q000201:enzymology*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment
D016623:Ion Pumps; D009097:Multienzyme Complexes; D000965:Antimony; D053501:Arsenite Transporting ATPases