The ways of realization of high specificity and efficiency of enteropeptidase.
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.
Mikhailova
Anna G
AG
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russian Federation. anna@enzyme.siobc.ras.ru
Likhareva
Viktoria V
VV
Teich
Niels
N
Rumsh
Lev D
LD
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004765:Enteropeptidase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D008954:Models, Biological; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014362:Trypsinogen / Q000378:metabolism
D050505:Mutant Proteins; D021122:Protein Subunits; D014362:Trypsinogen; D004765:Enteropeptidase