Quantitative Characterization of Three Carbonic Anhydrase Inhibitors by LESA Mass Spectrometry.
Quantitative Characterization of Three Carbonic Anhydrase Inhibitors by LESA Mass Spectrometry.
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DOI：
10.1021/jasms.2c00024
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发表时间：
2022-07-06
影响因子：
3.2
通讯作者：
Cooper HJ
中科院分区：
文献类型：
--
作者：
Illes-Toth E;Stubbs CJ;Sisley EK;Bellamy-Carter J;Simmonds AL;Mize TH;Styles IB;Goodwin RJA;Cooper HJ
液相萃取表面分析（LESA）结合天然质谱（MS），由于其灵敏度、速度和自动化，提供了独特的分析机会。在这里，我们研究这个工具是否可以用来定量探测蛋白质-配体相互作用，通过计算平衡解离常数（Kd值）。我们进行了本地LESA MS分析，包括牛碳酸酐酶II和配体氯噻嗪，丹磺酰胺，磺胺的一个良好的特征系统，并与直接输注质谱和表面等离子体共振测量获得的结果进行了比较。考虑了两种LESA方法：在一种方法中，蛋白质和配体在溶液中预混合，然后沉积并干燥到固体基底上进行LESA采样，在第二种方法中，仅将蛋白质干燥到基底上，并将配体包含在LESA中。采样溶剂。当蛋白质和配体预混时，发现直接输注MS和LESA MS获得的Kd值之间具有良好的一致性;然而，当配体在取样溶剂中时，从LESA MS测量确定的Kd值不一致。我们的研究结果表明，LESA MS是一个合适的工具，用于定量分析蛋白质-配体相互作用时，干燥的样品包括蛋白质和配体。
Liquid extraction surface analysis (LESA) coupled to native mass spectrometry (MS) presents unique analytical opportunities due to its sensitivity, speed, and automation. Here, we examine whether this tool can be used to quantitatively probe protein–ligand interactions through calculation of equilibrium dissociation constants (Kd values). We performed native LESA MS analyses for a well-characterized system comprising bovine carbonic anhydrase II and the ligands chlorothiazide, dansylamide, and sulfanilamide, and compared the results with those obtained from direct infusion mass spectrometry and surface plasmon resonance measurements. Two LESA approaches were considered: In one approach, the protein and ligand were premixed in solution before being deposited and dried onto a solid substrate for LESA sampling, and in the second, the protein alone was dried onto the substrate and the ligand was included in the LESA sampling solvent. Good agreement was found between the Kd values derived from direct infusion MS and LESA MS when the protein and ligand were premixed; however, Kd values determined from LESA MS measurements where the ligand was in the sampling solvent were inconsistent. Our results suggest that LESA MS is a suitable tool for quantitative analysis of protein–ligand interactions when the dried sample comprises both protein and ligand.
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影响因子：
7.4
作者：
El-Hawiet, Amr;Kitova, Elena N.;Klassen, John S.
通讯作者：
Klassen, John S.
影响因子：
7.4
作者：
Illes-Toth, Eva;Cooper, Helen J.
通讯作者：
Cooper, Helen J.
DOI：
10.1098/rspb.1976.0034
发表时间：
1976-01-01
期刊：
PROCEEDINGS OF THE ROYAL SOCIETY SERIES B-BIOLOGICAL SCIENCES
影响因子：
--
作者：
KING, RW;BURGEN, ASV
通讯作者：
BURGEN, ASV
影响因子：
8
作者：
Day, YSN;Baird, CL;Myszka, DG
通讯作者：
Myszka, DG
影响因子：
2.9
作者：
Cannon, MJ;Papalia, GA;Myszka, DG
通讯作者：
Myszka, DG