Lateral packing of the pancreatic lipase cofactor, colipase, with phosphatidylcholine and substrates.
The interaction of the pancreatic lipase cofactor colipase with a diacylphosphatidylcholine, acylglycerols, and free fatty acid was investigated by monitoring its adsorption to monomolecular lipid films. Surface pressure and colipase surface concentration were measured as a function of the initial lipid concentration and composition. Colipase adsorbs to a level of 28-30 pmol/cm2 to form a close-packed monolayer of protein and interacts strongly with all lipids when the lipid chain:colipase ratio is </=3. Consideration of the size difference between the protein and acyl groups suggests that in this regime the lipid is occupying the voids between tightly packed protein molecules. At lipid chain:colipase ratios >3, the triacylglycerol is excluded from the monolayer phase. Phosphatidylcholine, diacylglycerols, and free fatty acid remain in the monolayer phase up to </=25 lipid chain:colipase ratios. Geometrically over this range of compositions, the colipase molecules should be separated by up to 0-2 acyl chains. At higher lipid chain:colipase ratios, diacylglycerols are likely excluded from the monolayer phase. Anomalous behavior is observed with the fatty acid which at lipid chain:colipase ratios >25 induces higher levels of colipase adsorption than at lower ratios. This suggests the formation of a novel structure involving fatty acid and/or colipase. Phosphatidylcholine also remains in the interface at lipid chain:colipase ratios >3 but shows little additional interaction with colipase. However, fluorescence microscopy suggests that the phosphatidylcholine and colipase are miscible in the interface. The specificity demonstrated in this study suggests that colipase may regulate the type of surfaces to which colipase and, hence, lipase bind and may control the species distribution of substrate to which bound lipase is exposed.
Momsen
M M
MM
The Hormel Institute, University of Minnesota, Austin, Minnesota 55912, USA.
Dahim
M
M
Brockman
H L
HL
D001896:Boron Compounds; D003089:Colipases / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence; D010179:Pancreas / Q000201:enzymology*; D010713:Phosphatidylcholines / Q000737:chemistry*; D013379:Substrate Specificity
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D003089:Colipases; D005456:Fluorescent Dyes; D010713:Phosphatidylcholines; C027891:1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine