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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (03): 25-29    DOI:
    
Application of Histidine kinase (YycG) in screening inhibitor of Streptococcus pneumoniae
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Abstract  

Objective: To obtain a purified protein fragment with ATP-dependent kinase activity of the kinase domain of histidine kinase YycG in Streptococcus pneumoniae by a prokaryotic expression system and make use of its activity to search for new bacterial inhibitors. Methods: The protein of YycG kinase domain was expressed by a prokaryotic expression, identified by SDS-PAGE and Western Blot, and then purified by His-tagged Ni affinity column, and finally the activity of this purified protein was determined by a Kinase-Glo Luminescent Kinase Assay; Effective inhibitors were screened out from 105 candidate compounds based on their inhibitory action on the kinase activity of YycG protein, and were testified for their antibacterial effects in vitro. Results: The interest protein expressed in protocaryon was about 35KDa, reaching a purity of 95%, with obvious kinase activity of hydrolyzing ATP. Several effective kinase inhibitors were filtered out, and a selective one was verified overt bactericidal effects in vitro assay. Conclusion: The kinase inhibitors screened out based on their action on YycG may be important cues for developing antibacterial drugs or disinfectant.



Key wordshistidine kinase;purified protein;enzymatic activity;screening in vitro     
Received: 27 October 2008      Published: 31 March 2009
ZTFLH:  Q591  
Cite this article:

LI Na- Wang-Fei- Niu-Ci-Jiang- Wang-Gong- Li-Wei-Jiang- Yin-Nan-Lin- Zhang-Xue-Mei- Shu-Wei-Liang- Yin-Yi-Bing- Xu-Wen-Chun. Application of Histidine kinase (YycG) in screening inhibitor of Streptococcus pneumoniae. China Biotechnology, 2009, 29(03): 25-29.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I03/25

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