Cloning, expression, crystallization and preliminary X-ray crystallographic analysis of a human condensin SMC2 hinge domain with short coiled coils.
In higher eukaryotes, the condensin complex, which mainly consists of two structural maintenance of chromosomes (SMC) subunits, SMC2 (CAP-E) and SMC4 (CAP-C), plays a critical role in the formation of higher order chromosome structures during mitosis. Biochemical and electron-microscopic studies have revealed that the SMC2 and SMC4 subunits dimerize through the interaction of their hinge domains, forming a characteristic V-shaped heterodimer. However, the details of their function are still not fully understood owing to a lack of structural information at the atomic level. In this study, the human SMC2 hinge domain with short coiled coils was cloned, expressed, purified and crystallized in the orthorhombic space group C222 in native and SeMet-derivatized forms. Because of the poor diffraction properties of these crystals, the mutant Leu68-->SeMet was designed and crystallized in order to obtain the experimental phases. The SeMet-derivatized crystals of the mutant belonged to space group P3(2)12, with unit-cell parameters a=b=128.8, c=91.4 A. The diffraction data obtained from a crystal that diffracted to 2.4 A resolution were suitable for SAD phasing.
Kawahara
Kazuki
K
Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
Nakamura
Shota
S
Katsu
Yasuhiro
Y
Motooka
Daisuke
D
Hosokawa
Yuki
Y
Kojima
Yukiko
Y
Matsukawa
Keiko
K
Takinowaki
Hiroto
H
Uchiyama
Susumu
S
Kobayashi
Yuji
Y
Fukui
Kiichi
K
Ohkubo
Tadayasu
T
D000251:Adenosine Triphosphatases / Q000737:chemistry*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry*; D015870:Gene Expression; D006801:Humans; D046912:Multiprotein Complexes / Q000737:chemistry*; D017433:Protein Structure, Secondary
D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; C108312:condensin complexes; D000251:Adenosine Triphosphatases
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