The Ways of Realization of High Specificity and Efficiency of Enteropeptidase

Abstract

Comparative substrate analysis of full-length bovine enteropeptidase and trypsin, bovine and human enteropeptidase light chains was performed using model N-terminal dodecapeptides corresponding to wild-type human trypsinogen and pancreatitis-associated mutant trypsinogens K23R and D22G. The substitution of Lys residue by Arg at P1 leads to 2-fold increase in the efficiency of enteropeptidase hydrolysis; the absence of the negatively charged residue at P2 reduces the efficiency of such hydrolysis by two orders of magnitude. The difference in efficiency of peptide chain hydrolysis after Lys/Arg residues by enteropeptidase compared to trypsin is equal to the difference in hydrolysis by serine proteases of different primary specificity of their specific substrates.

Self‐degrading, MRI‐detectable hydrogel sensors with picomolar target sensitivity

“…Here, enterokinase was our target because it is specific to the trypsinogen‐trypsin conversion. Nonetheless, the mechanism for activating trypsin is well understood (26), making it possible to genetically or chemically modify the activation domain of trypsinogen or the catalytic activity of enterokinase (27). There are other identified autoactivating zymogens that are proteolytic (28, 29).…”

Heterogeneous catalysis on the phage surface: Display of active human enteropeptidase

“…For example, the catalytic constants of L-HEP for small peptide GD 4 K-na and mammalian trypsinogen activation peptide APFD 4 KIVGG are 10 and 17 times higher, respectively, than those of bovine enzyme [23,26]. Moreover, L-HEP and L-BEP also differ significantly in specificity toward various noncanonical peptides [15].…”

Human enteropeptidase light chain: Bioengineering of recombinants and kinetic investigations of structure and function

“…18,22,23 The monomeric rbEP L from E. coli proved superior to the native heterodimeric bEP at cleaving fusion proteins. 18 Human EP L has greater catalytic efficiency compared to bovine EP L for D 4 R$ X and D 4 K$ X substrates 24,25 and recombinant hEP light chain (rhEP L ) was shown to have a 10-fold higher specificity constant (k cat /K M ) for the D 4 K$ X sequence than bEP L .…”