Expression, purification and crystallization of Swi5 and the Swi5-Sfr1 complex from fission yeast.
The assembly of the presynaptic filament of recombinases represents the most important step in homologous recombination. The formation of the filament requires assistance from mediator proteins. Swi5 and Sfr1 have been identified as mediators in fission yeast and these proteins form a complex that stimulates strand exchange. Here, the expression, purification and crystallization of Swi5 and its complex with an N-terminally truncated form of Sfr1 (DeltaN180Sfr1) are presented. Analytical ultracentrifugation of the purified samples showed that Swi5 and the protein complex exist as tetramers and heterodimers in solution, respectively. Swi5 was crystallized in two forms belonging to space groups C2 and R3 and the crystals diffracted to 2.7 A resolution. Swi5-DeltaN180Sfr1 was crystallized in space group P2(1)2(1)2 and the crystals diffracted to 2.3 A resolution. The crystals of Swi5 and Swi5-DeltaN180Sfr1 are likely to contain one tetramer and two heterodimers in the asymmetric unit, respectively.
Kuwabara
Naoyuki
N
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Hashimoto
Hiroshi
H
Yamada
Noriyo
N
Unzai
Satoru
S
Ikeguchi
Mitsunori
M
Sato
Mamoru
M
Murayama
Yasuto
Y
Iwasaki
Hiroshi
H
Shimizu
Toshiyuki
T
D003460:Crystallization; D015870:Gene Expression; D011485:Protein Binding; D012568:Schizosaccharomyces / Q000737:chemistry*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D029702:Schizosaccharomyces pombe Proteins; C514357:Sfr1 protein, S pombe; C483769:Swi5 protein, S pombe
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