Physical and chemical characterization of the oligomerization state of the Aeromonas hydrophila lipase/acyltransferase.
Aeromonas glycerophospholipid:cholesterol acyl transferase undergoes a conformational transition upon activation by treatment with trypsin. Chemical cross-linking and sedimentation velocity analysis showed that the lipase dimerizes due to removal of a region near its C-terminus. The lipase monomer has a sedimentation coefficient s20.w = 2.83 S, whereas the dimer has s20.w = 3.65 +/- 0.22 S. Hydrodynamic analysis using these sedimentation values and the masses determined by mass spectrometry indicated that the monomers are aligned side-by-side in the dimer. An important change occurs in the apparent partial specific volume of the molecule upon activation.
Ausio
J
J
Department of Biochemistry and Microbiology, University of Victoria, BC, Canada.
van der Goot
F G
FG
Buckley
J T
JT
D000217:Acyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D016980:Aeromonas hydrophila / Q000201:enzymology*; D003432:Cross-Linking Reagents; D004120:Dimethyl Suberimidate; D004591:Electrophoresis, Polyacrylamide Gel; D008049:Lipase / Q000737:chemistry* / Q000302:isolation & purification; D046911:Macromolecular Substances; D013058:Mass Spectrometry; D008970:Molecular Weight; D014461:Ultracentrifugation
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D004120:Dimethyl Suberimidate; D000217:Acyltransferases; C020597:glycerophospholipid-cholesterol acyltransferase; D008049:Lipase