Recombinant Burkholderia pseudomallei Large-conductance mechanosensitive channel (mscL)

Introduction to Recombinant Burkholderia pseudomallei Large-conductance Mechanosensitive Channel (mscL)

The Recombinant Burkholderia pseudomallei Large-conductance mechanosensitive channel (mscL) is a protein derived from the bacterium Burkholderia pseudomallei, which is the causative agent of melioidosis, a severe infectious disease prevalent in tropical regions. Mechanosensitive channels like mscL are crucial for bacterial survival under osmotic stress by allowing the efflux of ions and small molecules to maintain cellular integrity.

Structure and Function of mscL

• Structure: The mscL protein is a pentameric structure composed of five identical subunits. Each subunit contains two transmembrane helices (TM1 and TM2) connected by a periplasmic loop. The channel's pore is formed by the TM1 helices, which are highly conserved across different species .

• Function: mscL channels are activated by membrane tension, allowing them to open and release ions and small molecules from the cell. This function is essential for maintaining cellular osmotic balance and preventing lysis under conditions of rapid osmotic changes .

Recombinant Production and Characteristics

• Production: Recombinant Burkholderia pseudomallei mscL is produced using molecular biology techniques, where the gene encoding mscL is cloned into an expression vector and expressed in a suitable host organism. This allows for large-scale production of the protein for research and potential therapeutic applications .

• Characteristics: The recombinant mscL protein is typically stored in a Tris-based buffer with 50% glycerol to maintain stability. It is recommended to store the protein at -20°C or -80°C to prevent degradation. Working aliquots can be stored at 4°C for up to one week .

Research Findings and Applications

• Biological Significance: Understanding the structure and function of mscL in Burkholderia pseudomallei can provide insights into the pathogen's survival mechanisms and potential targets for therapeutic intervention. Research on mscL may also contribute to the development of novel antimicrobial strategies .

• Potential Applications: The study of mechanosensitive channels like mscL can inform the design of new drugs or treatments targeting bacterial osmotic regulation. Additionally, recombinant mscL proteins can be used as tools in biotechnology for developing sensors or devices sensitive to mechanical stress .