Engineering MerR for Sequestration and MerA for Reduction of Toxic Metals and Radionuclides

Summers, Anne O.

Univ. of Georgia Research Foundation

关键词: Calorimetry;    Titration Mercury, Methylmercury,Biotransformation, Horizontal Gene Transfer;    Crystallography;    Dialysis;    Gold;   

DOI  :  10.2172/944097
RP-ID  :  DOE/ER/62865-3
RP-ID  :  FG02-99ER62865
RP-ID  :  944097

美国|英语

来源: UNT Digital Library

【 摘 要 】

The objectives of this project were (1) to alter a metalloregulatory protein (MerR) so that it would bind other toxic metals or radionuclides with similar affinity so that the engineered protein itself and/or bacteria expressing it could be deployed in the environment to specifically sequester such metals and (2) to alter the mercuric reductase, MerA, to reduce radionuclides and render them less mobile. Both projects had a basic science component. In the first case, such information about MerR illuminates how proteins discriminate very similar metals/elements. In the second case, information about MerA reveals the criteria for transmission of reducing equivalents from NADPH to redox-active metals. The work involved genetic engineering of all or parts of both proteins and examination of their resultant properties both in vivo and in vitro, the latter with biochemical and biophysical tools including equilibrium and non-equilibrium dialysis, XAFS, NMR, x-ray crystallography, and titration calorimetry. We defined the basis for metal specificity in MerR, devised a bacterial strain that sequesters Hg while growing, characterized gold reduction by MerA and the role of the metallochaperone domain of MerA, and determined the 3-D structure of MerB, the organomercurial lyase.