Structural and evolutionary relationships in lipase mechanism and activation.
Lipases that break down triglycerides to monoglycerides and glycerol are characterised by low or no activity in water; in the presence of an oil/water interface, however, their activity increases markedly. The structural and chemical basis for this phenomenon, referred to as interfacial activation, has been revealed by the crystal structures of a fungal lipase and a human pancreatic lipase which evidently have a divergent evolutionary history. These studies reveal that: (1) In both enzymes the catalytic sidechains are Asp:His:Ser, the same as occur in the serine proteases. The active atoms on this catalytic triad have essentially identical stereochemistry in the serine proteases and in these two lipases. The amino acids themselves, however, have quite different conformations and orientations. (2) In both enzymes the catalytic groups are buried and inaccessible to the surrounding solvent. Burial in these two lipases is brought about by a small stretch of helix (the lid) which sits over the active site. (3) In both enzymes this helical lid presents non-polar sidechains over the catalytic group, and polar sidechains to the enzyme surface. Although the 'lids' are very similar in construction in the two enzymes, they belong to very different parts of the polypeptide chain. (4) Although the amino acid sequences have no identity (except at the active serine) the two enzymes show a similar architectural framework consisting of a central five-stranded parallel beta sheet structure. The catalytic groups decorate this beta sheet structure in a strikingly similar way though there are also some significant differences. The crystal structure of the complex between the fungal enzyme and a substrate analogue demonstrates how the helical lid is displaced to reveal the active site. The movement of the lid also greatly enlarges the non-polar surface at the active surfaces and buries previously exposed polar residues. The movement of the lid also helps to create the appropriate movement at the oxyanion hole. It is possible to define the stereochemistry at the active site and to identify the positioning of the fatty acid and the glycerol moieties.
Dodson
G G
GG
Department of Chemistry, University of York, Heslington, UK.
Lawson
D M
DM
Winkler
F K
FK
D000595:Amino Acid Sequence; D001665:Binding Sites; D002384:Catalysis; D004789:Enzyme Activation; D006801:Humans; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship
D008049:Lipase