Examination of Protein Phosphorylation in Gram-Positive Bacteria Biofilm

Abstract

All gram-positive bacteria, such as Staphylococcus aureus, produce biofilms as a method to protect themselves from their surroundings; however, these communities of bacteria can be particularly harmful to the human body. Biofilms are able to form on medical devices inserted into bodies, and bacteria can travel from primary infection sites to produce secondary infections throughout the body. Uncontrolled biofilms can cause sepsis and eventually death. Further, treating infectious biofilms is challenging because they produce a protective coating that allows the bacteria to evade immune system responses and treatments with antibiotics. To stop these infections early in their paths, the mechanisms behind biofilm formation must be fully understood, and target proteins in these pathways must be inhibited. The following study can be used to help identify the proteins involved in the formation of biofilms by studying how the phosphorylation of proteins changes upon biofilm formation. Staphylococcus aureus bacteria cells were lysed to extract protein, and protein concentrations were quantized with a bicinchoninic acid (BCA) assay. Proteins were separated by molecular weight with 1D SDS-PAGE. The fluorescent Pro-Q Diamond Phosphoprotein Gel Stain was used in conjunction with SYPRO Ruby dye to demonstrate protein phosphorylation. This study is currently ongoing, and research methods and techniques are being optimized.