Methodology of electrospray ion trap mass spectrometry for analyzing the non-covalent binding of protein and low-molecular-weight ligand / 药学学报
Acta Pharmaceutica Sinica
; (12): 408-412, 2007.
Article
em Zh
| WPRIM
| ID: wpr-281883
Biblioteca responsável:
WPRO
ABSTRACT
A new MS-titration method for the non-covalent binding of protein-ligand based on the research of berberine and alpha1-acid glycoprotein was established. The major presumption of new method is that the total concentration of protein-ligand complex is approximately the same as the total concentration of acting protein if a certain extent of affinity is existed between protein and ligand, in addition, the mole amount of acting ligand is more than that of acting protein. The non-covalent binding behaviours between berberine and alpha1-acid glycoprotein was studied by using electrospray ionization ion trap mass spectrometry (ESI-ITMS) , and the results were verified by fluorescence quenching method. The results showed that the binding behaviours between berberine and alpha1-acid glycoprotein, for example, stability constant, number of binding site, type of the main binding force, were almost the same by using the new MS-titration method and fluorescence quenching method. Comparing with the reported MS-titration method, the presented MS-titration method in this paper is more simple and applicable, does not demand much for the devices, and can lead to reliable results in same cases.
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Base de dados:
WPRIM
Assunto principal:
Ligação Proteica
/
Berberina
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Orosomucoide
/
Proteínas
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Espectrometria de Massas por Ionização por Electrospray
/
Mapeamento de Interação de Proteínas
/
Ligantes
/
Metabolismo
/
Métodos
Idioma:
Zh
Revista:
Acta Pharmaceutica Sinica
Ano de publicação:
2007
Tipo de documento:
Article