Hi guys, I am doing a research related the role of bile acids in protease activation. When I add hydrophobic bile acid or bile salt to trypsin, its activity measured by Fluorometer (RFU) decrease in concentration depending manner. It seems that protein denaturation plays a role here. But how to prove it and how to prevent it? Thank you!
Dear Trung!
Could You look at this article:
ROLE OF BILE ACIDS IN THE ACTIVATION OF TRYPSINOGEN BY ENTEROKINASE: DISTURBANCE OF TRYPSINOGEN ACTIVATION IN PATIENTS WITH INTRAHEPATIC BILIARY ATRESIA
https://www.gastrojournal.org/article/S0016-5085(74)80043-0/pdf
Dear Alexandr, thank you very much for prompt reply with a nice paper. Looking at it, I learnt more that certain hile salts like NaT, GDCA... can enhance the role of enterokinase in activating trysinogen. In my own experiment, I also add bile acid (taurolithocholic acid and deoxycholic acid) directly to trypsinogen with or without enterokinase ans saw that bile acid itself even can cleave the trysinogen after 3 hours whivh is shown by lower band in Westernblot. However the point here is i other where when we add bile acid directly to pure trypsin, we see that its activivity reduce in concentration depending maner. There are may be also the direct interaction between Trypsin and bile acids but I have not known how to prove this.
Isothermal titration calorimetry (ITC) would measure the heat of reaction between bile acid and protein, surface plasmon resonance (SPR) would be another option.
Dear Dr Buxbaum, thank you very much for your answer! Engelbert Buxbaum
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