Dithiothreitol

Dithiothreitol

Dithiothreitol chemical structure

Dithiothreitol (DTT) is a commonly used reducing agent in biochemistry and molecular biology. It is a small molecule with the chemical formula C4H10O2S2 and a molecular weight of 154.25 g/mol. DTT is known for its ability to break disulfide bonds, which are important in maintaining the structure and function of proteins.

History

DTT was first synthesized in the 1950s by William J. Lushbaugh and colleagues at the University of Cincinnati. They discovered its reducing properties while studying the effects of radiation on biological systems. Since then, DTT has become a widely used tool in various research fields.

Structure and Properties

DTT consists of a central carbon atom bonded to two thiol (-SH) groups and two hydroxyl (-OH) groups. The thiol groups are responsible for its reducing properties. DTT is a white crystalline solid that is soluble in water and most organic solvents. It has a melting point of approximately 42-44°C.

Applications

Protein Research

DTT is commonly used in protein research to reduce disulfide bonds. Disulfide bonds play a crucial role in stabilizing the three-dimensional structure of proteins. By breaking these bonds, DTT can denature proteins, allowing for further analysis or manipulation. DTT is often used in protein purification, protein folding studies, and protein structure determination.

Polymerase Chain Reaction (PCR)

DTT is also used in PCR, a widely used technique in molecular biology. PCR requires the use of heat-stable DNA polymerases, which can be inhibited by disulfide bonds. DTT is added to the PCR reaction mixture to reduce any disulfide bonds present, thus preventing the inhibition of DNA polymerase activity.

Safety Considerations

DTT is a strong reducing agent and should be handled with care. It can cause skin and eye irritation, and prolonged exposure may lead to respiratory sensitization. It is important to wear appropriate protective equipment, such as gloves and goggles, when working with DTT. Additionally, DTT should be stored in a cool, dry place away from oxidizing agents.

See Also

• Disulfide bond
• Protein purification
• Polymerase Chain Reaction