Determining the affinity of anti-mitotic compounds binding to colchicine binding site of tubulin by affinity probe capillary electrophoresis

doi:10.1016/j.jchromb.2019.05.010

	Determining the affinity of anti-mitotic compounds binding to colchicine binding site of tubulin by affinity probe capillary electrophoresis
	Xu, Mei 1; Xu, Yao 1,2; Shen, Yuting 1,2; Lou, Chunli1,3 ; Zheng, Mengmeng 1,2; Kang, Jingwu1,3
	2019-07-15
发表期刊	JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES (IF:2.8[JCR-2023],2.8[5-Year])
ISSN	1570-0232
卷号	1121 页码:66-71
发表状态	已发表
DOI	10.1016/j.jchromb.2019.05.010
摘要	The colchicine binding site of tubulin is often used to screen the anti-mitotic compounds, which are widely used as anti-cancer therapies. In the present work, an affinity probe capillary electrophoresis (APCE) method was developed for determining the affinity of anti-mitotic compounds. To this end, a fluorescently labeled affinity probe, 5-carboxyfluorescein-colchicine (F-colchicine), was prepared for the affinity competition experiment. The probe can form a stable complex with tubulin with the binding stoichiometry of 0.75, and the dissociation constant K-d of the complex was determined as 5.7 x 10(-5) mol/L. In the affinity competition experiment, F-colchicine was incubated with tubulin and the test compound in the solution. The F-colchicine-tubulin complexes and free F-colchicine were quickly separated by CE and the concentration of free F-colchicine was accurately determined with the laser induced fluorescence detection. The affinity constant of the tested compound can be measured with the affinity competition binding curve. The enantiomers of the anti-mitotic compound were evaluated by using the method. The binding affinity of the enantiomers displayed an enantioselective manner. Compared to other affinity binding assay methods, our method is more straightforward, more accurate, and more cost-effective.
关键词	Affinity probe capillary electrophoresis Anti-mitotic compounds Colchicine binding site Tubulin
收录类别	SCI ; SCIE ; EI
语种	英语
资助项目	Strategic Priority Research Program of the Chinese Academy of Sciences[XDB20020200]
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS类目	Biochemical Research Methods ; Chemistry, Analytical
WOS记录号	WOS:000471207200008
出版者	ELSEVIER SCIENCE BV
EI入藏号	20192106955197
EI主题词	Binding energy ; Capillary electrophoresis ; Cost effectiveness ; Dissociation ; Enantiomers
EI分类号	Chemistry:801 ; Chemical Reactions:802.2 ; Industrial Economics:911.2
WOS关键词	BIOLOGICAL EVALUATION ; FLUORESCENT-PROBE ; INHIBITORS ; DESIGN ; ASSAY ; CONSTANTS ; EXTRACTS ; ANALOGS
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/48963
专题	物质科学与技术学院_硕士生物质科学与技术学院_特聘教授组_康经武组
通讯作者	Kang, Jingwu
作者单位	1.Chinese Acad Sci, Ctr Excellence Mol Synth, Shanghai Inst Organ Chem, State Key Lab Bioorgan & Nat Prod Chem, Lingling Rd 345, Shanghai 200032, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.ShanghaiTech Univ, Sch Phys Sci & Technol, Haike Rd 100, Shanghai 200120, Peoples R China
通讯作者单位	物质科学与技术学院
推荐引用方式 GB/T 7714	Xu, Mei,Xu, Yao,Shen, Yuting,et al. Determining the affinity of anti-mitotic compounds binding to colchicine binding site of tubulin by affinity probe capillary electrophoresis[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2019,1121:66-71.
APA	Xu, Mei,Xu, Yao,Shen, Yuting,Lou, Chunli,Zheng, Mengmeng,&Kang, Jingwu.(2019).Determining the affinity of anti-mitotic compounds binding to colchicine binding site of tubulin by affinity probe capillary electrophoresis.JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,1121,66-71.
MLA	Xu, Mei,et al."Determining the affinity of anti-mitotic compounds binding to colchicine binding site of tubulin by affinity probe capillary electrophoresis".JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 1121(2019):66-71.