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Study on the Function and Crystal Structures of YdiV and YdaM in E.coli Signaling Pathway Transduced by C-di-GMP

Author: GuoLiMing
Tutor: GuLiChuan
School: Shandong University
Course: Microbiology
Keywords: Quorum sensing effect C - di - GMP YdiV YdaM
CLC: Q78
Type: Master's thesis
Year: 2010
Downloads: 194
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Abstract


Cluster of bacterial cells in cell-to-cell communication, quorum sensing effect. Quorum sensing effect to regulate a variety of physiological processes of the bacteria, including pathogens movement and pathogenicity. Quorum sensing with automatic inducer (AI) for signaling molecules to achieve the communication between cells. AI is an extracellular signaling molecules, eventually leading to cell physiological and biochemical reactions by means of intracellular signaling molecules, C-di-GMP is a second messenger molecules. C-di-GMP is a widely present in bacteria of the second messenger molecules, the regulation of a variety of physiological activities of the bacterial cells, for example, the pathogenicity of Vibrio cholerae, Pseudomonas aeruginosa biofilm formation of E. coli biological the film formation, and so on. C-di-GMP concentration in the process of signal transmission, the changes will affect its effects as. C-di-GMP concentration responsible for the catalytic synthesis of double guanylate cyclase and is responsible for the catalytic hydrolysis of phosphodiesterase to adjust. C-di-GMP, the regulation mechanism of the C-di-GMP concentration level directly affects Vibrio cholerae, Pseudomonas aeruginosa, Salmonella pathogenicity. Regulation of the level of C-di-GMP concentration to inhibit the function of the protein, or directly regulate the function of the C-di-GMP has important guiding significance for the prevention and control of the disease and treatment. Accurate screening with a C-di-GMP-related protein, accurate positioning of the active site of the protein, to elucidate biochemical reaction mechanism, is the use of a C-di-GMP for pharmaceutical therapeutic targets need to be resolved. In this paper, based on the existing conditions of the laboratory, the use of E. coli as carrier bacteria to study the structure and function of homologous proteins, respectively EAL domain-containing YdiV protein containing GGDEF domain YdaM protein structure at all levels, attempts to resolve the crystal structure of YdiV the preliminary crystallographic part of the data, and try to by the SAD method parse out YdiV the crystal structure. Protein overexpression with related gene ydiv, the ydam knockout inspection YdiV YdaM biochemical functions within the bacterial cells, to test its correlation with C-di-GMP concentration level, and the formation of biofilm. Reference the in Known Good EAL / GGDEF play activity conditions, YdiV YdaM no activity assay enzyme activity by extracellular intracellular experiments significant biological activity detected, indicating that the C-di-GMP concentration adjusting C-di-GMP regulation of biofilm there is a wide range of regulatory networks.

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