Investigation of human copper transporter proteins pore formation through number and brightness analysis

Abstract

In this study we present the investigation of pore formation of human copper transporter (hCtr) proteins in live cancer cells using the number and brightness (N&B) analysis. Copper is one of the most important metals for cell metabolism, considered acrucial cofactor for several processes in eukaryotic cells such as respiration, detoxification andsecretion1. Any imbalance on copper homeostasis can lead to severe diseases. In this homeostasis process, the hCtrs are the proteins responsible for copper uptake by the cell. It is known that hCtrs can exists as monomers, dimers or trimers and that the trimers seem to be responsible for the functional pore formation2. The N&B analysis is a potent statistical method based on fluorescent intensity fluctuations of single pixels recorded using a photon-counting confocal microscopy3. Our results indicate that after stimulation with a cooper chelator the hCtrs aggregate. We were able to map the aggregation of hCtr monomers by using N&B. In order to understand the pore formation in cancer cells, we will treat the cells with different concentrations of copper