Activation of nonspecific lipase (EC 3.1.1.-) by bile salts.
The enzyme nonspecific lipase (EC 3.1.1.-) from rat pancreas has been isolated and its amino acid composition determined. The amino acid composition confirms more indirect evidence that nonspecific lipase is not the same enzyme as cholesteryl ester hydrolase. Activation of the enzymatic activity by bile salts has been studied by equilibrium dialysis, gel filtration, light scattering, circular dichroism and fluorescence polarization. The binding of bile salt by the enzyme is saturable and is associated with a conformational change. Upon binding cholate, the protein experiences a decrease in beta-structure with no significant change in alpha-helix content, an increase in apparent Stokes radius, a decrease in light scattering properties, and a slight decrease in polarization of the intrinsic tryptophan fluorescence. Attachment of bile salt is associated with decreased reactivity of essential sulfhydryl groups, but no detectable change in reactivity of amino groups. A change to a more nearly spherical shape upon binding bile salt would be consistent with the experimental observations, but the exact sites of binding remain uncertain.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001647:Bile Acids and Salts / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D019826:Cholic Acid; D002793:Cholic Acids / Q000494:pharmacology; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003840:Deoxycholic Acid / Q000378:metabolism / Q000494:pharmacology; D003956:Dialysis; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D005454:Fluorescence Polarization; D008027:Light; D008049:Lipase / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D008823:Micelles; D010179:Pancreas / Q000201:enzymology*; D010795:Phthalic Acids / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D012542:Scattering, Radiation; D013656:Taurocholic Acid / Q000494:pharmacology
D000596:Amino Acids; D001647:Bile Acids and Salts; D002793:Cholic Acids; D008823:Micelles; D010795:Phthalic Acids; D003840:Deoxycholic Acid; C024629:dimethyl phthalate; D013656:Taurocholic Acid; D008049:Lipase; D019826:Cholic Acid