Part:BBa_K3843000

PEA-binding salmon trypsin (1UTM)

Trypsin is a ubiquitous protease in vertebrates due to its central role in digestion. Trypsin cleaves peptide bonds occurring after positively charged amino acids, such as lysine and arginine. 1UTM (pictured below) is a trypsin from Salmo salar (Atlantic salmon). It has a variety of known inhibitors, typically biogenic amines (Leiros et al., 2009). These noncompetitive inhibitors bind to allosteric sites, thereby inactivating trypsin's proteolytic function. Of particular interest to Waterloo iGEM 2021 was the ability of phenylethylamine (PEA, pictured below in green), a dopamine precursor, to act as an inhibitor of 1UTM, with a dissociation constant Kd = 0.000972 (Leiros et al., 2009; RCSB PDB, n.d.). This would allow for 1UTM to be used as a PEA-binding protein. Detection and quantification of phenylethylamine, using 1UTM fused to horseradish peroxidase in a microfluidic flow assay, would give insight into an individual's probability of having ADHD.

3D structure of 1UTM, as per the crystal structure of 1UTM on the PDB. 1UTM is coloured in orange, while phenylethylamine (PEA) is coloured in green. The 3D structure was developed using UCSF Chimera.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]