Pressure effects on catalysis by alkaline phosphatase reconstituted in lipid bilayers.
Bovine intestinal alkaline phosphatase (EC 3.1.3.1) was reconstituted into lipid bilayers by a dilution method using n-octylglucopyranoside. From the kinetic measurements at various pressures, the volume of activation (delta V not equal to) and volume change in substrate binding (delta V) were estimated for free and reconstituted ALP. The delta V not equal to and delta V values for free ALP and reconstituted ALP in the gel state liposome showed opposite tendencies (-23 ml . mol-1 [delta V not equal to], 35 ml . mol-1 [delta V] for free ALP and 27 ml . mol-1 [delta V not equal to], -36 ml . mol-1 [delta V] for reconstituted ALP, respectively), which suggest both strong desolvation effect of enzyme molecule by the surrounding lipids and drastic conformational change of the enzyme molecule by the reconstitution into liposomes.
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D004134:Dimyristoylphosphatidylcholine; D007422:Intestines / Q000201:enzymology; D007700:Kinetics; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D011312:Pressure; D013816:Thermodynamics
D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D000469:Alkaline Phosphatase; D004134:Dimyristoylphosphatidylcholine