Design and application of chemical probes targeting hydrolytic protein families

来源 :第11届海峡两岸细胞生物学学术研讨会 | 被引量 : 0次 | 上传用户:meiyajun1008
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  Chemical approaches have been used in analyzing biological systems such as enzyme inhibitors/activators, substrates, molecular probes, and protein modifiers.With proper designed chemical compounds,the function of proteins could be controlled, modified, monitored, or manipulated.This chemistry-driven approach can be used to answer biological questions that cannot be easily addressed by traditional biochemical or genetic approaches.A chemical probe consists of a properly positioned reactive group which forms a covalent bond with the target protein and a reporter for visualization and/or purification of the covalently bound target.The reactive group and the reporter are separated by a linker so that neither reaction of the reactive group with enzymatic active sites nor affinity recognition/detection of the reporter is sterically hindered by the other part of the probe.We have adopted a mechanism-based approach to develop activity probes for hydrolases such as tyrosine phosphatases and glycosidases.We have also designed active-site direct probes to target kinases.Here I will report our current progress in developing chemical probes targeting serine hydrolases.Members of serine hydrolyase family are crucial for the regulation of numerous cellular events, including cell growth, tissue differentiation, neoplastic formation, as well as tumor metastasis.However, due to the lack of a proper tool that enables the analysis of them, the current characterization of serine hydrolases preceded at a slower pace.Applying fluorophosphonate as the reactive group, we have synthesized and characterized a series of benzyl fluorophosphonate-based probes for labeling serine hydrolases.These newly designed probes target to serine hydrolases with high specificity.Moreover,through altering the hydrophobicity, several of the probes could enter cells to label serine hydrolases in vivo.We anticipate that our chemical probes will provide excellent technical platforms in proteomic research,serine hydrolases activity analysis, and facilitate the process of finding important disease-related targets.
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