Talin-1 head(hereinafter referred to as TH)is the head structure of talin protein,which contains a four-point-one-protein/ezrin/radixin/moesin(FERM)domain.Its F1 domain contains an unstructured loop of 30 amino acids(...Talin-1 head(hereinafter referred to as TH)is the head structure of talin protein,which contains a four-point-one-protein/ezrin/radixin/moesin(FERM)domain.Its F1 domain contains an unstructured loop of 30 amino acids(139-168),which does not interact with other domains.Because TH doesn’t get the crystal structure and whether the unstructured loop has obvious influence on the TH secondary structure,therefore,the truncated talin-1 headΔ139-168(hereinafter referred to as THΔ)was constructed and its structure and the impact of stability after truncation were analyzed.Molecular biology and structural biology methods were used to construct prokaryotic expression vectors of TH and THΔ,explore and optimize the expression conditions of recombinants,and they were purified by affinity chromatography and FPLC gel filtration chromatography.Finally,a large number of stable,high-purity protein samples were prepared successfully.The physicochemical properties and structural stability of the proteins were investigated by dynamic light scattering and circular dichroism.The results showed that the THΔsecondary structure of the truncated body did not change significantly,the structural stability was enhanced and the resistance to guanidine hydrochloride and high temperature was stronger.展开更多
With our interest in （S）-3-aminopyrrolidine derivatives,we further screened inhibition activities of three hit compounds on many other kinases with the results demonstrating that this series of compounds shows bette...With our interest in （S）-3-aminopyrrolidine derivatives,we further screened inhibition activities of three hit compounds on many other kinases with the results demonstrating that this series of compounds shows better anticancer activities,which might result from the main block of PI3K/Akt signaling pathway and the inhibition of Abelson murine leukemia viral oncogene homolog（ABL） kinase,as well as some epidermal growth factors.Further structure modification demonstrates that benzylsulfonyl group is the necessary functional group contributing to the biological activity that will be helpful to guiding us to optimize these （S）-3-aminopyrrolidine derivatives.展开更多
Abstract CC chemokine receptor 4 （CCR4） is a kind of G-protein-coupled receptor, which plays a pivotal role in allergic inflammation. The interaction between 2-（2-（4-chloro-phenyl）-5-{[（naphthalen-1- ylmethyl）-...Abstract CC chemokine receptor 4 （CCR4） is a kind of G-protein-coupled receptor, which plays a pivotal role in allergic inflammation. The interaction between 2-（2-（4-chloro-phenyl）-5-{[（naphthalen-1- ylmethyl）-carbamlyl]-methyl-4-oxo-thiazolidin-3-yl）-N-（3-morpholin-4-yl-propyi）-acetamide （S009） and the N-terminal extracellular tail （ML40） of CCR4 has been validated to be high affinity by capillary zone electrophoresis （CZE）.The S009 is a known CCR4 antagonist. Now, a series of new thiourea derivatives have been synthesized. Compared with positive control S009, they were screened using ML40 as target by CZE to find some new drugs for allergic inflammation diseases. The synthesized compounds XJH-5, XJH-4, XJH-17 and XJH-1 displayed the interaction with ML40, but XJH-9, XJH-10, XJH-I 1, XJH-12, XJH-13, XJH-14, XJH-3, XJH-8, XJH-6, XJH-7, XJH-15, XJH-16 and XJH-2 did not bind to ML40. Both qualification and quantification characterizations of the binding were determined. The affinity of the four compounds was valued by the binding constant, which was similar with the results of chemotactic experiments. The established CEZ method is capable of sensitive and fast screening for a series of lactam analogs in the drug discovery for allergic inflammation diseases.展开更多
基金Supported by the Scientific Research Personnel of Northeast Agricultural University(518001)。
文摘Talin-1 head(hereinafter referred to as TH)is the head structure of talin protein,which contains a four-point-one-protein/ezrin/radixin/moesin(FERM)domain.Its F1 domain contains an unstructured loop of 30 amino acids(139-168),which does not interact with other domains.Because TH doesn’t get the crystal structure and whether the unstructured loop has obvious influence on the TH secondary structure,therefore,the truncated talin-1 headΔ139-168(hereinafter referred to as THΔ)was constructed and its structure and the impact of stability after truncation were analyzed.Molecular biology and structural biology methods were used to construct prokaryotic expression vectors of TH and THΔ,explore and optimize the expression conditions of recombinants,and they were purified by affinity chromatography and FPLC gel filtration chromatography.Finally,a large number of stable,high-purity protein samples were prepared successfully.The physicochemical properties and structural stability of the proteins were investigated by dynamic light scattering and circular dichroism.The results showed that the THΔsecondary structure of the truncated body did not change significantly,the structural stability was enhanced and the resistance to guanidine hydrochloride and high temperature was stronger.
文摘With our interest in （S）-3-aminopyrrolidine derivatives,we further screened inhibition activities of three hit compounds on many other kinases with the results demonstrating that this series of compounds shows better anticancer activities,which might result from the main block of PI3K/Akt signaling pathway and the inhibition of Abelson murine leukemia viral oncogene homolog（ABL） kinase,as well as some epidermal growth factors.Further structure modification demonstrates that benzylsulfonyl group is the necessary functional group contributing to the biological activity that will be helpful to guiding us to optimize these （S）-3-aminopyrrolidine derivatives.
基金supported by the National Key New Drug Creation Program of China (2009ZX09103-724)the National Natural Science Foundation of China grants(30872292,90813025 and 81072612)+2 种基金the Natural Science Foundation of Beijing (7102107)the Open Foundation of State Key Laboratory of Natural and Biomimetic Drugs(K20090207)the National New Drug Research and Development Project of China (2009ZX09301-010)
文摘Abstract CC chemokine receptor 4 （CCR4） is a kind of G-protein-coupled receptor, which plays a pivotal role in allergic inflammation. The interaction between 2-（2-（4-chloro-phenyl）-5-{[（naphthalen-1- ylmethyl）-carbamlyl]-methyl-4-oxo-thiazolidin-3-yl）-N-（3-morpholin-4-yl-propyi）-acetamide （S009） and the N-terminal extracellular tail （ML40） of CCR4 has been validated to be high affinity by capillary zone electrophoresis （CZE）.The S009 is a known CCR4 antagonist. Now, a series of new thiourea derivatives have been synthesized. Compared with positive control S009, they were screened using ML40 as target by CZE to find some new drugs for allergic inflammation diseases. The synthesized compounds XJH-5, XJH-4, XJH-17 and XJH-1 displayed the interaction with ML40, but XJH-9, XJH-10, XJH-I 1, XJH-12, XJH-13, XJH-14, XJH-3, XJH-8, XJH-6, XJH-7, XJH-15, XJH-16 and XJH-2 did not bind to ML40. Both qualification and quantification characterizations of the binding were determined. The affinity of the four compounds was valued by the binding constant, which was similar with the results of chemotactic experiments. The established CEZ method is capable of sensitive and fast screening for a series of lactam analogs in the drug discovery for allergic inflammation diseases.
