Deleterious effect of Brij 35 on alkyl 2-pyrones and other hydrophobic inhibitors of human sputum and leucocyte elastase.
Brij 35 significantly reduced the inhibitory activity of hydrophobic alkyl 2-pyrones, oleic acid and alkyl peptides towards human sputum and leucocyte elastase, whereas 4-methoxy-6-(2'-hydroxy-2'-(carbobutyloxy)-vinyl)-2-pyrone, alpha-1-proteinase inhibitor and a sulfated chitosan were unaffected. The effect of Brij 35 on elastase appeared to be irreversible, since dialysis against Brij-free buffer was not accompanied by a return to inhibitory activity by the first group of inhibitors. However, passage through an ionic-exchange column was effective in removing the detergent from the enzyme. Brij 35 is also an activator of the elastases: kcat for Boc-Ala-4-nitrophenyl ester and methylsuccinyl-Ala-Ala-Pro-Val-4-nitroanilide increased by 20% and 40%, respectively in the presence of 0.015% Brij 35. Binding of the substrates to the enzyme is unaffected, since Km is unchanged.
Cook
L
L
Chemistry Department, La Trobe University, Bundoora, Australia.
Ternai
B
B
D003902:Detergents / Q000009:adverse effects*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors*; D000077423:Polidocanol; D011092:Polyethylene Glycols / Q000009:adverse effects*; D013501:Surface-Active Agents / Q000009:adverse effects*
D003902:Detergents; D013501:Surface-Active Agents; D000077423:Polidocanol; D011092:Polyethylene Glycols; D010196:Pancreatic Elastase