Crystal structure of a supercharged variant of the human enteropeptidase light chain

Proteins. 2012 Jul;80(7):1907-10. doi: 10.1002/prot.24084. Epub 2012 May 10.


The highly specific serine protease human enteropeptidase light chain cleaves the Asp4Lys recognition sequence and represents an interesting enzyme for biotechnological applications. The human enzyme shows 10 times faster kinetics compared to other animal sources but low solubility under low salt conditions, which hampers protein production and crystallization. Therefore, a supercharged variant (N6D/G21D/G22D/N142D/K210E/C112S) with increased solubility was used for crystallization. The structure (resolution, 1.9 Å) displays a typical α/β trypsin-like serine protease-fold. The mutations introduced for protein supercharging generate larger clusters of negative potential on both sites of the active cleft but do not affect the structural integrity of the protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Enteropeptidase / chemistry*
  • Enteropeptidase / genetics
  • Enzyme Stability
  • Humans
  • Kinetics
  • Models, Molecular
  • Mutation
  • Protein Conformation
  • Protein Subunits / chemistry
  • Static Electricity
  • X-Ray Diffraction


  • Protein Subunits
  • Enteropeptidase