Revisiting the hydrolysis of ampicillin catalyzed by Temoneira-1 β-lactamase, and the effect of Ni(II), Cd(II) and Hg(II)
Abstract ?-Lactamases grant resistance to bacteria against ?-lactam antibiotics. The active center of TEM-1 ?-lactamase accommodates a Ser-Xaa-Xaa-Lys motif. TEM-1 ?-lactamase is not a metalloenzyme but it possesses several putative metal ion binding sites. The sites composed of His residue pairs ch...
Elmentve itt :
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| Dokumentumtípus: | Cikk |
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2023
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| Sorozat: | PROTEIN SCIENCE
32 No. 12 |
| Tárgyszavak: | |
| doi: | 10.1002/pro.4809 |
| mtmt: | 34393301 |
| Online Access: | http://publicatio.bibl.u-szeged.hu/31947 |
| Tartalmi kivonat: | Abstract ?-Lactamases grant resistance to bacteria against ?-lactam antibiotics. The active center of TEM-1 ?-lactamase accommodates a Ser-Xaa-Xaa-Lys motif. TEM-1 ?-lactamase is not a metalloenzyme but it possesses several putative metal ion binding sites. The sites composed of His residue pairs chelate borderline transition metal ions such as Ni(II). In addition, there are many sulfur-containing donor groups that can coordinate soft metal ions such as Hg(II). Cd(II) may bind to both types of the above listed donor groups. No significant change was observed in the circular dichroism spectra of TEM-1 ?-lactamase on increasing the metal ion content of the samples, with the exception of Hg(II) inducing a small change in the secondary structure of the protein. A weak nonspecific binding of Hg(II) was proven by mass spectrometry and 119mHg perturbed angular correlation spectroscopy. The hydrolytic process of ampicillin catalyzed by TEM-1 ?-lactamase was described by the kinetic analysis of the set of full catalytic progress curves, where the slow, yet observable conversion of the primary reaction product into a second one, identified as ampilloic acid by mass spectrometry, needed also to be considered in the applied model. Ni(II) and Cd(II) slightly promoted the catalytic activity of the enzyme while Hg(II) exerted a noticeable inhibitory effect. Hg(II) and Ni(II), applied at 10??M concentration, inhibited the growth of E. coli BL21(DE3) in M9 minimal medium in the absence of ampicillin, but addition of the antibiotic could neutralize this toxic effect by complexing the metal ions. |
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| Terjedelem/Fizikai jellemzők: | 20 |
| ISSN: | 0961-8368 |