| Exposure to high levels of arsenic causes a wide range of health effects. Arsenic binding to the sulfhydryl group of cysteine in proteins, affecting protein's function, is one of the possible mechanisms. This thesis focuses on understanding arsenic binding to proteins and developing affinity capture techniques for the analysis of arsenic binding proteins.;The binding of arsenicals with p53 and PARP-1 was studied using HPLC-ICP-MS, affinity capture and Western blot analysis. Weak binding of p53 and PARP-1 to phenylarsine oxide was observed.;An arsenic affinity technique was developed to capture the specific arsenic binding proteins. The captured proteins were subsequently identified by mass spectrometry. Over seventy proteins in A549 cells were captured and identified. Using the available structural information of the proteins, the possible cysteines binding to arsenic were proposed. This technique was further applied to identify proteins in the nuclear extract of A549 cells treated with either benzo[α]pyrene diol expoxide alone or in combination with arsenite. |
