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| Expression, Purification and Crystal Growth Studies on Thiamin Momophosphate Kinase (ThiL) from Shigella flexneri 2a Strain 301 |
| SHANG Gui-jun1, DING Zhi-qiang1, ZHAO Zi-hua1, NIE Rong-xin2, GAO Wei1, CANG Huai-xing2 |
1. Beijing Forestry University, Beijing 100083, China;
2. Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China |
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Abstract Thiamin monophosphate kinase (ThiL) catalyzes the ATP-dependent phosphorylation of thiamin monophosphate (TMP) to form thiamin pyrophosphate (TPP), the active form of vitamin B1. ThiL is a member of a small ATP binding superfamily. The gene of ThiL from Shigella flexneri 2a(strain 301)was constructed into the expression vector and over expressed in the E.coli. Then it was purified through two steps of chromatography leading to the high purity of the protein. The purified protein was screened for crystallization. The hit condition was optimized and gave rise to the single crystals for X-ray diffraction, which is the fundamental step for its structure determination, illumination of its catalytic mechanism, and corresponding drug design.
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Received: 06 September 2010
Published: 25 December 2010
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