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Deleterious effect of Brij 35 on alkyl 2-pyrones and other hydrophobic inhibitors of human sputum and leucocyte elastase

Cook, L.; Ternai, B.

Biochemistry International 17(4): 637-646

1988

ISSN/ISBN: 0158-5231
PMID: 3240314
Document Number: 319132
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.

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