Plasmodium falciparum rab6 GTPase: expression, purification, crystallization and preliminary crystallographic studies.
The Plasmodium falciparum rab6 gene encodes a 208 amino-acid polypeptide. Two recombinant versions of P. falciparum Rab6 protein were expressed in Escherichia coli: the full-length protein and a truncated form containing residues 1-175. Both forms were purified from the soluble fraction of bacterial extract and were purified by ion-exchange chromatography and size-exclusion chromatography. Purified proteins were crystallized at pH 6.5 using the hanging-drop vapor-diffusion technique at room temperature. The full-length protein diffracted to 2.4 A and belongs to the tetragonal space group P4(3)2(1)2 or P4(1)2(1)2, with unit-cell parameters a = b = 80. 6, c = 90.4 A. The crystals of the truncated protein were isomorphous with those of the full-length construct and diffracted X-rays to 2.2 A resolution.
Chattopadhyay
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Division of Geographic Medicine, School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA. debasish@polaris.cmc.uab.edu
Smith
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CD
Barchue
J
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Langsley
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D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D017125:Genes, Protozoan; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020691:rab GTP-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
C094135:Rab6 protein; D011994:Recombinant Proteins; D020691:rab GTP-Binding Proteins