Phenylmethylsulfonyl fluoride

Phenylmethylsulfonyl fluoride (PMSF) is an organosulfur compound with the molecular formula C₇H₇FO₂S. It is a serine protease inhibitor widely used in biochemical and molecular biology research to prevent protein degradation during sample preparation. PMSF is a potent and irreversible inhibitor of serine proteases such as trypsin, chymotrypsin, thrombin, and subtilisin, as it covalently modifies the active site serine residue in these enzymes.

Chemical and Physical Properties

• Molecular Formula: C₇H₇FO₂S
• Molecular Weight: 174.19 g/mol
• Appearance: White to off-white crystalline solid
• Melting Point: Approximately 92–96°C
• Solubility: PMSF is soluble in organic solvents such as ethanol, methanol, isopropanol, acetone, and DMSO but poorly soluble in water.
• Stability: PMSF is hydrolyzed in aqueous solutions, especially at alkaline pH, which limits its effectiveness over time in buffer solutions.

Mechanism of Action

PMSF functions as an irreversible inhibitor of serine proteases by covalently binding to the hydroxyl (-OH) group of the active site serine residue. Phenylmethylsulfonyl fluoride reaction leads to enzyme inactivation, preventing protein degradation by endogenous or exogenous proteases. The inhibition occurs rapidly in solutions with neutral to slightly basic pH but is less effective in highly acidic or strongly basic conditions. Buy Phenylmethylsulfonyl fluoride in Finland

Applications in Research and Industry

1. Protease Inhibition in Protein Purification

   • PMSF is commonly used to protect proteins from degradation during extraction, purification, and storage.
   • It is added to lysis buffers and extraction solutions to maintain protein integrity.

2. Biochemical and Molecular Biology Research

   • Used to study the structure and function of proteases.
   • Helps in investigating protein-protein interactions by preventing unwanted proteolysis.

3. Cell Biology and Immunology

   • Used in cell lysates and tissue homogenates to prevent proteolytic degradation of cellular components.
   • Used in immunoprecipitation and Western blotting protocols to preserve proteins.

4. Pharmaceutical and Drug Development

   • PMSF serves as a reference inhibitor in studies evaluating new protease inhibitors for drug discovery.
   • It helps in enzyme kinetics studies and understanding protease activity.

Handling, Safety, and Storage

• Toxicity: PMSF is highly toxic and poses serious health risks upon inhalation, ingestion, or skin absorption. It is classified as a hazardous chemical with potential effects on the respiratory system and nervous system.
• Irritant Properties: It can cause severe irritation to the skin, eyes, and mucous membranes. Proper personal protective equipment (PPE) such as gloves, lab coats, and safety goggles should be worn when handling PMSF.
• Storage: PMSF should be stored in a cool, dry place, away from moisture and direct light. It is typically stored as a dry powder at -20°C and dissolved in an anhydrous solvent before use to prevent hydrolysis.
• Disposal: Due to its toxicity, PMSF should be disposed of according to local hazardous waste disposal regulations.