An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins
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. H...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-07-01
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| Series: | Results in Chemistry |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221171562500356X |
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| Summary: | 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. |
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| ISSN: | 2211-7156 |