Trypsin

Chemistry and function

Trypsin is secreted into the intestine, where it acts to hydrolyse proteins into smaller pH of about 8 and optimal operating temperature of about 37°C.^[1]

The polypeptides.

Trypsin is produced in the autocatalysis), so only a small amount of enterokinase is necessary to start the reaction. This activation mechanism is common for most serine proteases, and serves to prevent autodigestion of the pancreas.

The activity of trypsins is not affected by the inhibitor tosyl phenylalanyl chloromethyl ketone TPCK, which deactivates mass spectrometry, the specificity of cleavage is important.

Involvement in disease

One consequence of inheriting the autosomal recessive disease cystic fibrosis is a deficiency of trypsin and other digestive enzymes from the pancreas. This leads to the disorder termed meconium ileus. This disorder involves intestinal obstruction (ileus) due to overly thick meconium which is normally broken down by trypsins and other proteases, then passed in feces.

Storage

Trypsins should be stored at very cold temperatures (between −20°C and −80°C) to prevent autolysis (self-cleavage). Autolysis may also be prevented by storage of trypsins at pH 3 or by using trypsin modified by e.g. reductive methylation. When the pH is adjusted back to pH 8 activity returns.

Applications

Trypsin is available in high quantities in pancreases, and can be purified rather easily. Hence it has been used widely in various biotechnological processes.

In a tissue culture lab, trypsins are used to re-suspend cells adherent to the cell culture dish wall during the process of harvesting cells.

Trypsin can also be used to dissociate dissected cells. For example, prior to cell fixing and sorting.

Trypsins can be used to breakdown casein in breast milk. If trypsin is added to a solution of milk powder, the breakdown of casein will cause the milk to become translucent. The rate of reaction can be measured by using the amount of time it takes for the milk to turn translucent.

Trypsin is commonly used in biological research during proteomics experiments to digest proteins into peptides for mass spectrometry analysis. Trypsin is particularly suited for this, since it has a very well defined specificity.

Trypsin can also be used to dissolve blood clots in its microbial form and treat inflammation in its pancreatic form.

Trypsin is used in baby food to pre-digest it. It can break down the protein molecules which helps the baby to digest it as its stomach is not strong enough to digest bigger protein molecules.

See also

• Trypsin inhibitor

References

1. ^a  Promega PDF - 58kB

• IUBMB entry for 3.4.21.4
• BRENDA references for 3.4.21.4 (Recommended.)
• PubMed references for 3.4.21.4
• PubMed Central references for 3.4.21.4
• Google Scholar references for 3.4.21.4

• IUBMB entry for 3.4.21.4

• KEGG entry for 3.4.21.4

• BRENDA entry for 3.4.21.4

• NiceZyme view of 3.4.21.4

• EC2PDB: PDB structures for 3.4.21.4

• PRIAM entry for 3.4.21.4

• PUMA2 entry for 3.4.21.4

• IntEnz: Integrated Enzyme entry for 3.4.21.4

• MetaCyc entry for 3.4.21.4

• Atomic-resolution structures of enzymes belonging to this class

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