Work

An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins

Year: 2025

Type: article

Abstract: Superbug infection caused by New Delhi metallo-β-lactamase (NDM-1) has grown into an emerging threat. EDTA is one of the most commonly used Zn(II) clelators for preparation of apo-metallo-β-lactamases (apo-MβLs) and further Co(II)-substituted MβLs for spectroscopic characterization of the enzymes. However, the removal of EDTA from the apo-proteins is rarely verified. Here, we provide a convenient and quick isothermal titration calorimetry (ITC) method for real-time monitoring of the removal of EDTA from apo-NDM-1. This ITC method can be used for monitoring the removal of clelators from apo-MβLs, even wide apo-matalloproteins, which enable one to prepare Co(II)-substituted proteins for further structural and mechanistic studies.

Source: Results in Chemistry

DOI: 10.1016/j.rechem.2025.102373

Authors: Ying Ge, Wen-Ming Wang, Tingjun Liu

Institutions: Xijing University, Northwest University, PLA Air Force Xi'an Flying College

Language: English

Topic: thermodynamics and calorimetric analyses

Subfield: Physical and Theoretical Chemistry

Field: Chemistry

Domain: Physical Sciences

Open Access status: gold

Estimated APC paid: $1,870

Funders: Innovative Research Group Project of the National Natural Science Foundation of China, Dalian High-Level Talent Innovation Program, Xijing University, Natural Science Foundation of Shaanxi Provincial Department of Education