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. 1995 Jan;4(1):44–57. doi: 10.1002/pro.5560040107

Crystallographic study of the structure of colipase and of the interaction with pancreatic lipase.

M P Egloff 1, L Sarda 1, R Verger 1, C Cambillau 1, H van Tilbeurgh 1
PMCID: PMC2142970  PMID: 7773176

Abstract

Colipase (Mr 10 kDa) confers catalytic activity to pancreatic lipase under physiological conditions (high bile salt concentrations). Previously determined 3-A-resolution X-ray structures of lipase-colipase complexes have shown that, in the absence of substrate, colipase binds to the noncatalytic C-terminal domain of pancreatic lipase (van Tilbeurgh H, Sarda L, Verger R, Cambillau C, 1992, Nature 359:159-162; van Tilbeurgh et al., 1993a, Nature 362:814-820). Upon lipid binding, conformational changes at the active site of pancreatic lipase bring a surface loop (the lid) in contact with colipase, creating a second binding site for this cofactor. Covalent inhibition of the pancreatic lipase by a phosphonate inhibitor yields better diffracting crystals of the lipase-colipase complex. From the 2.4-A-resolution structure of this complex, we give an accurate description of the colipase. It confirms the previous proposed disulfide connections (van Tilbeurgh H, Sarda L, Verger R, Cambillau C, 1992, Nature 359:159-162; van Tilbeurgh et al., 1993a, Nature 362:814-820) that were in disagreement with the biochemical assignment (Chaillan C, Kerfelec B, Foglizzo E, Chapus C, 1992, Biochem Biophys Res Commun 184:206-211). Colipase lacks well-defined secondary structure elements. This small protein seems to be stabilized mainly by an extended network of five disulfide bridges that runs throughout the flatly shaped molecule, reticulating its four finger-like loops. The colipase surface can be divided into a rather hydrophilic part, interacting with lipase, and a more hydrophobic part, formed by the tips of the fingers. The interaction between colipase and the C-terminal domain of lipase is stabilized by eight hydrogen bonds and about 80 van der Waals contacts. Upon opening of the lid, three more hydrogen bonds and about 28 van der Waals contacts are added, explaining the higher apparent affinity in the presence of a lipid/water interface. The tips of the fingers are very mobile and constitute the lipid interaction surface. Two detergent molecules that interact with colipase were observed in the crystal, covering part of the hydrophobic surface.

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Selected References

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  1. Adler M., Lazarus R. A., Dennis M. S., Wagner G. Solution structure of kistrin, a potent platelet aggregation inhibitor and GP IIb-IIIa antagonist. Science. 1991 Jul 26;253(5018):445–448. doi: 10.1126/science.1862345. [DOI] [PubMed] [Google Scholar]
  2. Borgström B., Wieloch T., Erlanson-Albertsson C. Evidence for a pancreatic pro-colipase and its activation by trypsin. FEBS Lett. 1979 Dec 15;108(2):407–410. doi: 10.1016/0014-5793(79)80574-8. [DOI] [PubMed] [Google Scholar]
  3. Brünger A. T., Kuriyan J., Karplus M. Crystallographic R factor refinement by molecular dynamics. Science. 1987 Jan 23;235(4787):458–460. doi: 10.1126/science.235.4787.458. [DOI] [PubMed] [Google Scholar]
  4. Chaillan C., Kerfelec B., Foglizzo E., Chapus C. Direct involvement of the C-terminal extremity of pancreatic lipase (403-449) in colipase binding. Biochem Biophys Res Commun. 1992 Apr 15;184(1):206–211. doi: 10.1016/0006-291x(92)91179-t. [DOI] [PubMed] [Google Scholar]
  5. Cozzone P. J., Canioni P., Sarda L., Kaptein R. 360-MHz nuclear magnetic resonance and laser photochemically induced dynamic nuclear polarization studies of bile salt interaction with porcine colipase A. Eur J Biochem. 1981;114(1):119–126. doi: 10.1111/j.1432-1033.1981.tb06181.x. [DOI] [PubMed] [Google Scholar]
  6. De Caro A., Figarella C., Amic J., Michel R., Guy O. Human pancreatic lipase: a glycoprotein. Biochim Biophys Acta. 1977 Feb 22;490(2):411–419. doi: 10.1016/0005-2795(77)90016-2. [DOI] [PubMed] [Google Scholar]
  7. Erlanson-Albertsson C., Larsson A. A possible physiological function of pancreatic pro-colipase activation peptide in appetite regulation. Biochimie. 1988 Sep;70(9):1245–1250. doi: 10.1016/0300-9084(88)90191-5. [DOI] [PubMed] [Google Scholar]
  8. Erlanson-Albertsson C., Larsson A. Importance of the N-terminal sequence in porcine pancreatic colipase. Biochim Biophys Acta. 1981 Aug 24;665(2):250–255. doi: 10.1016/0005-2760(81)90009-6. [DOI] [PubMed] [Google Scholar]
  9. Erlanson-Albertsson C. Pancreatic colipase. Structural and physiological aspects. Biochim Biophys Acta. 1992 Apr 8;1125(1):1–7. doi: 10.1016/0005-2760(92)90147-n. [DOI] [PubMed] [Google Scholar]
  10. Erlanson-Albertsson C. The existence of pro-colipase in pancreatic juice. Biochim Biophys Acta. 1981 Nov 23;666(2):299–300. doi: 10.1016/0005-2760(81)90121-1. [DOI] [PubMed] [Google Scholar]
  11. Erlanson C., Charles M., Astier M., Desnuelle P. The primary structure of porcine colipase II. II. The disulfide bridges. Biochim Biophys Acta. 1974 Jul 7;359(1):198–203. doi: 10.1016/0005-2795(74)90143-3. [DOI] [PubMed] [Google Scholar]
  12. Fukuoka S., Taniguchi Y., Kitagawa Y., Scheele G. Full length cDNA sequence encoding canine pancreatic colipase. Nucleic Acids Res. 1990 Sep 25;18(18):5549–5549. doi: 10.1093/nar/18.18.5549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grasberger B. L., Clore G. M., Gronenborn A. M. High-resolution structure of Ascaris trypsin inhibitor in solution: direct evidence for a pH-induced conformational transition in the reactive site. Structure. 1994 Jul 15;2(7):669–678. doi: 10.1016/s0969-2126(00)00067-8. [DOI] [PubMed] [Google Scholar]
  14. Huang K., Strynadka N. C., Bernard V. D., Peanasky R. J., James M. N. The molecular structure of the complex of Ascaris chymotrypsin/elastase inhibitor with porcine elastase. Structure. 1994 Jul 15;2(7):679–689. doi: 10.1016/s0969-2126(00)00068-x. [DOI] [PubMed] [Google Scholar]
  15. Janin J., Chothia C. The structure of protein-protein recognition sites. J Biol Chem. 1990 Sep 25;265(27):16027–16030. [PubMed] [Google Scholar]
  16. Kabsch W., Sander C. Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features. Biopolymers. 1983 Dec;22(12):2577–2637. doi: 10.1002/bip.360221211. [DOI] [PubMed] [Google Scholar]
  17. McIntyre J. C., Schroeder F., Behnke W. D. Synthesis and characterization of the dansyltyrosine derivatives of porcine pancreatic colipase. Biochemistry. 1990 Feb 27;29(8):2092–2101. doi: 10.1021/bi00460a019. [DOI] [PubMed] [Google Scholar]
  18. Rathelot J., Canioni P., Bosc-Bierne I., Sarda L., Kamoun A., Kaptein R., Cozzone P. J. Limited trypsinolysis of porcine and equine colipases. Spectroscopic and kinetic studies. Biochim Biophys Acta. 1981 Dec 29;671(2):155–163. doi: 10.1016/0005-2795(81)90129-x. [DOI] [PubMed] [Google Scholar]
  19. Rathelot J., Julien R., Bosc-Bierne I., Gargouri Y., Canioni P., Sarda L. Horse pancreatic lipase. Interaction with colipase from various species. Biochimie. 1981 Mar;63(3):227–234. doi: 10.1016/s0300-9084(81)80196-4. [DOI] [PubMed] [Google Scholar]
  20. Renaud W., Dagorn J. C. cDNA sequence and deduced amino acid sequence of human preprocolipase. Pancreas. 1991 Mar;6(2):157–161. doi: 10.1097/00006676-199103000-00005. [DOI] [PubMed] [Google Scholar]
  21. Rydel T. J., Tulinsky A., Bode W., Huber R. Refined structure of the hirudin-thrombin complex. J Mol Biol. 1991 Sep 20;221(2):583–601. doi: 10.1016/0022-2836(91)80074-5. [DOI] [PubMed] [Google Scholar]
  22. Saudek V., Atkinson R. A., Pelton J. T. Three-dimensional structure of echistatin, the smallest active RGD protein. Biochemistry. 1991 Jul 30;30(30):7369–7372. doi: 10.1021/bi00244a003. [DOI] [PubMed] [Google Scholar]
  23. Sternby B., Erlanson-Albertsson C. Measurement of the binding of human colipase to human lipase and lipase substrates. Biochim Biophys Acta. 1982 Apr 15;711(1):193–195. doi: 10.1016/0005-2760(82)90025-x. [DOI] [PubMed] [Google Scholar]
  24. van Tilbeurgh H., Gargouri Y., Dezan C., Egloff M. P., Nésa M. P., Ruganie N., Sarda L., Verger R., Cambillau C. Crystallization of pancreatic procolipase and of its complex with pancreatic lipase. J Mol Biol. 1993 Jan 20;229(2):552–554. doi: 10.1006/jmbi.1993.1054. [DOI] [PubMed] [Google Scholar]
  25. van Tilbeurgh H., Sarda L., Verger R., Cambillau C. Structure of the pancreatic lipase-procolipase complex. Nature. 1992 Sep 10;359(6391):159–162. doi: 10.1038/359159a0. [DOI] [PubMed] [Google Scholar]

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