Specific one-stage method for assay of enterokinase activity by release of radiolabelled activation peptides from alpha-N-[3H]acetyl-trypsinogen and the effect of calcium ions on the enzyme activity.
We report a novel assay method for enterokinase capable of detecting approx. 1 fmol of enzyme. The method depends on quantification of the release of specifically radiolabelled activation peptides from bovine trypsinogen and is unaffected by trypsin inhibitors. The assay is applicable to biological fluids such as serum. The substrate was produced by selective epsilon-amidination of bovine trypsinogen followed by acetylation with [3H]acetic anhydride and deprotection. The assay has been used to study the effects of pH, Ca2+, ionic strength abd glycodeoxycholate on enterokinase activity.
D000107:Acetylation; D002118:Calcium / Q000494:pharmacology*; D010450:Endopeptidases / Q000378:metabolism*; D004765:Enteropeptidase / Q000378:metabolism*; D004789:Enzyme Activation; D006002:Glycodeoxycholic Acid / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008722:Methods; D009994:Osmolar Concentration; D010455:Peptides / Q000378:metabolism*; D014316:Tritium; D014362:Trypsinogen / Q000378:metabolism*
C032591:N-acetyl-trypsinogen; D010455:Peptides; D014316:Tritium; D006002:Glycodeoxycholic Acid; D014362:Trypsinogen; D010450:Endopeptidases; D004765:Enteropeptidase; D002118:Calcium