[Expression, purification and crystallization of rat brain kinesin].
Sheng Wu Gong Cheng Xue Bao
; 30(3): 485-91, 2014 Mar.
Article
em Zh
| MEDLINE
| ID: mdl-25007584
ABSTRACT
Kinesin is a motor protein that uses the energy from ATP hydrolysis to move along the microtubule system. To investigate how the chemical energy stored in ATP is converted to mechanical movement, the corresponding N-terminal region of rat brain kinesin was expressed in BL21-Codon Plus (DE3)-RP competent cells. After SP-cation exchange chromatography and size exclusion chromatography, the protein yield reached 10 mg/L culture with the purity above 95%. The purified protein had ATPase activity and specifically reacted with the kinesin antibody in the Western blotting analysis. The purified kinesin was crystallized under the following condition 1.7 mol/L (NH4)2SO4, 500 mmol/L NaCl, 20% glycerol. The kinesin crystal can diffract up to 2.0 angstroms resolution.
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MEDLINE
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Cinesinas
Limite:
Animals
Idioma:
Zh
Revista:
Sheng Wu Gong Cheng Xue Bao
Ano de publicação:
2014
Tipo de documento:
Article