1887

Abstract

MerR, the metalloregulator of the bacterial mercury resistance () operon, binds Hg(II) with high affinity. To study the mechanism of metal-induced activation, a small protein was previously engineered embodying in a single polypeptide the metal-binding domain (MBD) ordinarily formed between two monomers of MerR. Here the physiological and biochemical properties of MBD expressed on the cell surface or in the cytosol were examined, to better understand the environments in which specific metal binding can occur with this small derivative. Over 20 000 surface copies of MBD were expressed per cell, with metal stoichiometries of ∼1·0 Hg(II) per MBD monomer. Cells expressing MBD on their surface in rich medium bound 6·1-fold more Hg(II) than those not expressing MBD. Although in nature cells use the entire operon to detoxify mercury, it was interesting to note that cells expressing only MBD survived Hg(II) challenge and recovered more quickly than cells without MBD. Cell-surface-expressed MBD bound Hg(II) preferentially even in the presence of a 22-fold molar excess of Zn(II) and when exposed to equimolar Cd(II) in addition. MBD expressed in the cystosol also afforded improved survival from Hg(II) exposure for and for the completely unrelated bacterium .

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2006-03-01
2019-10-23
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vol. , part 3, pp. 709 - 719

[PDF](40 kb) encodes single-chain Fv of an anti-digoxin antibody fragment and was part of the vector as received. Intermediate plasmid pB30D-SalI was deleted in part of and of , which encodes the chloramphenicol acetyltransferase; encodes the β-lactamase. pASK-MBD was constructed by Jonathan Caguiat (Song , 2004). The name pSD-MBD denotes surface-displayed MB. Engineered single-chain, antiparallel, coiled coil mimics the MerR metal binding site. , 1861-1868.



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