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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Elsevier
2025-07-01
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| Series: | Results in Chemistry |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S221171562500356X |
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| author | Ying Ge Wen-Ming Wang Tingjun Liu |
| author_facet | Ying Ge Wen-Ming Wang Tingjun Liu |
| author_sort | Ying Ge |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-46b81b27e0894059a0b0ecfacfa2bcf9 |
| institution | Kabale University |
| issn | 2211-7156 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Results in Chemistry |
| spelling | doaj-art-46b81b27e0894059a0b0ecfacfa2bcf92025-08-20T04:00:33ZengElsevierResults in Chemistry2211-71562025-07-011610237310.1016/j.rechem.2025.102373An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteinsYing Ge0Wen-Ming Wang1Tingjun Liu2Biomedical Research Center of Xijing University, Medical College, Xijing University, Xi'an 710127, PR China; Corresponding author.Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University, PR ChinaAir Force Xi'an Flying College, Xi'an 710300, ChinaSuperbug 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.http://www.sciencedirect.com/science/article/pii/S221171562500356XMetallo-β-lactamasesIsothermal titration calorimetryApo-proteinEDTA monitoring |
| spellingShingle | Ying Ge Wen-Ming Wang Tingjun Liu An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins Results in Chemistry Metallo-β-lactamases Isothermal titration calorimetry Apo-protein EDTA monitoring |
| title | An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins |
| title_full | An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins |
| title_fullStr | An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins |
| title_full_unstemmed | An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins |
| title_short | An isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra-acetic acid from apo-proteins |
| title_sort | isothermal titration calorimetry method for monitoring the removal of ethylenediamine tetra acetic acid from apo proteins |
| topic | Metallo-β-lactamases Isothermal titration calorimetry Apo-protein EDTA monitoring |
| url | http://www.sciencedirect.com/science/article/pii/S221171562500356X |
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