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Method Development for Identification and Verification of Protein Metal‐Binding Sites
Author(s) -
Healy Kelly Lynn
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.631.21
Subject(s) - metallothionein , cadmium , binding site , biochemistry , chemistry , cysteine , zinc , metalloprotein , mutant , zinc finger , biology , gene , transcription factor , organic chemistry , enzyme
Cadmium is a nonessential heavy metal that is found in many common items such as batteries, pigments, wheat, and tobacco, and is known to cause nephrotoxicity, hepatotoxicity, and osteotoxicity. Furthermore, studies have demonstrated that cadmium exposure leads to an accumulation of mutations by disrupting the DNA repair activities of mismatch repair proteins and polynucleotide kinase, as well as by creating reactive oxygen species in the cell. In response to heavy metal exposure, organisms such as bacteria, plants, and animals express metallothionein (MT). MT is rich in cysteine residues, promoting binding to divalent metal cations such as zinc and cadmium. MT has a very slow evolution rate and the cysteine residues are conserved throughout species, indicating that it is essential for life, yet the exact residues where metals bind are unknown. Metal‐binding sites were identified by a three‐dimensional model of rabbit metallothionein‐1A built by predictive protein databases based on homologous structures. Metal‐binding fluorophores were used to quantify the amount of cadmium, zinc, magnesium, and manganese bound to different constructs of MT. Locating binding site mutations in MT is the first step to identifying mutant proteins and correcting function. In addition to identifying the metal‐binding sites in rabbit MT‐1, this project will allow us to use this methodology to locate binding sites in other proteins that are not structurally characterized as well as MT. Support or Funding Information This project was funded by St. Mary's Undergraduate Research Fellowship at St. Mary's College of Maryland and by the Cove Point Natural Heritage Trust's Ruth Mathes Scholarship. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .