A General Mass Spectrometry-Based Assay for the Quantitation of Protein-Ligand Binding Interactions in Solution

详细信息    查看全文

• 作者：Kendall D. Powell ; Sina Ghaemmaghami ; Michael Z. Wang ; Liyuan Ma ; Terrance G. Oas ; and Michael C. Fitzgerald
• 刊名：Journal of the American Chemical Society
• 出版年：2002
• 出版时间：September 4, 2002
• 年：2002
• 卷：124
• 期：35
• 页码：10256 - 10257
• 全文大小：42K
• 年卷期：v.124,no.35(September 4, 2002)
• ISSN：1520-5126

文摘

A new method that utilizes matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and exploits the hydrogen/deuterium (H/D) exchange properties of proteins was developed for measuring the thermodynamic properties of protein-ligand complexes in solution. Dissociation constants (K_d values) determined by the method for five model protein-ligand complexes that included those with small molecules, nucleic acids, peptides, and other proteins were generally in good agreement with K_d values measured by conventional methods. Important experimental advantages of the described method over existing methods include: the ability to make measurements in a high-throughput and automated fashion, the ability to make measurements using only picomole quantitities of protein, and the ability to analyze either purified or unpurified protein-ligand complexes.