Cathepsin D Crystal Structures and Lysosomal Sorting

  • Peter Metcalf
  • Martin Fusek
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 362)


The D is not for dull! In this chapter we discuss the features of cathepsin D which are interesting because of their relevance to studies on lysosomal targeting. We will not describe the active site cleft, the centre of attention for aspartic proteinases with greater specificity or with more obvious medical significance. Mechanisms of protein targeting are of fundamental interest, and increasingly implicated in medicine. Unlike the aspartic proteases with previously known structures, cathepsin D is intracellular, and we solved crystal structures for human spleen cathepsin D and pepstatin inhibited bovine liver cathepsin D partly to learn more about what special features of this molecule cause it to be targeted to lysosomes1–3. There is one other solved structure of a lysosomal enzyme; the cysteine proteinase cathepsin B4. We are currently comparing its surface with that of cathepsin D to search for expected common features, as discussed in the following paragraphs.


Lysosomal Enzyme Aspartic Protease Molecular Replacement Active Site Cleft Human Cathepsin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    F. Bieber, V. Brachvogel, R. Arni, M. Fusek and P. Metcalf, Crystallization and initial crystallographic results for pepstatin A inhibited bovine cathepsin D, J.Mol.Biol. 227, 1265–1268 (1992).PubMedCrossRefGoogle Scholar
  2. 2.
    M. Fusek, M. Baudys and P. Metcalf, Purification and crystallization of human cathepsin D, J.Mol.Biol. 226, 555–557 (1992).PubMedCrossRefGoogle Scholar
  3. 3.
    P. Metcalf and M. Fusek, Two crystal structures for cathepsin D: the lysosomal targeting signal and active site, EMBO J. 12, 1293–302 (1993).PubMedGoogle Scholar
  4. 4.
    D. Musil, D. Zucic, D. Turk, I. Mayr, R. A. Engh, R. Huber, T. Popovic, V. Turk, T. Towatari, N. Katunuma and W. Bode, The refined 2.15 Å X-ray crytal structure of human liver cathepsin B: the structural basis for its specificity., EMBO J. 10, 2321–2330 (1991).PubMedGoogle Scholar
  5. 5.
    Kornfeld, Structure and function of the mannose 6-phosphate/insulin like growth factor II receptors, Annu. Rev. Biochem. 61, 307–330 (1992).PubMedCrossRefGoogle Scholar
  6. 6.
    T. Ludwig, C. E. Ovitt, J. Remmler, P. Lobel, U. Ruther and B. Hoflack, Targeted disruption of the mouse cation-dependent mannose 6-phosphate receptor results in partial missorting of multiple lysosomal enzymes, EMBO J. 12, 5225–5235 (1993).PubMedGoogle Scholar
  7. 7.
    T. Ludwig, H. Munier-Lehmann, M. Hollinshead, C. Ovitt, P. Lobel and B. Hoflack, Differential sorting of lysosomal enzymes in mannose 6-phosphate receptor-deficient fibroplasts (submitted), (1994).Google Scholar
  8. 8.
    A. Köster, P. Saftig, U. Matzner, K. von Figura, C. Peters and R. Pohlmann, Targeted disruption of the Mr 46 000 mannose 6-phsophate receptor gene in mice results in misrouting of lysosomal proteins, EMBL J. 12, 5219–5223 (1993).Google Scholar
  9. 9.
    S. Kornfeld, Trafficking of lysosomal enzymes in normal and disease states, J. Clin. Invest. 77, 1–6 (1986).PubMedCrossRefGoogle Scholar
  10. 10.
    T. J. Baranski, P. L. Faust and S. Kornfeld, Generation of lysosomal enzyme targeting signal in the secretory protein pepsinogen, Cell 63, 281–291 (1990).PubMedCrossRefGoogle Scholar
  11. 11.
    P. Novick and P. Brennwald, Friends and Family: The role of Rab GTPases in vesicular traffic, Cell 75, 597–601 (1993).PubMedCrossRefGoogle Scholar
  12. 12.
    D. J. Kilonsky, L. M. Banta and S. D. Emr, Intracellular sorting and processing of a yeast vacuolar hydrolase: proteinase A propeptide contains vacuolar targeting information., Mol. Cell. Biol. 8, 2105–2116 (1988).Google Scholar
  13. 13.
    A. T. Brünger, Extension of molecular replacement: A new search strategy based on patterson correlation refinement, Acta Cryst. A46, 46–57 (1990).Google Scholar
  14. 14.
    A. T. Brünger, Free R Value: a novel statistical quantity for assessing the accuracy of crystal structures, Nature 355, 472–475 (1992).PubMedCrossRefGoogle Scholar
  15. 15.
    C. Abad-Zapatero, T. H. Rydel and J. Erickson, Revised 2.3Å structure of porcine pepsin: Evidence for a Flexible Subdomain, Proteins 8, 62–81 (1990).PubMedCrossRefGoogle Scholar
  16. 16.
    A. Sali, B. Veerapandian, J. Coper, D. Modd, T. Hofmann and T. Blundell, Domain flexibility in aspartic proteases, Proteins 12, 158–170 (1992).PubMedCrossRefGoogle Scholar
  17. 17.
    A. R. Sielecki, A. A. Federov, A. Boodhoo, N. S. Andreeva and M. N. G. James, Molecular and crystal structures of monoclinic porcine pepsin refined at 1.8Å resolution, J. Mol. Biol. 214, 143–170 (1990).PubMedCrossRefGoogle Scholar
  18. 18.
    S. J. Gould, G.-A. Keller, M. Schneider, S. H. Howell, L. J. Garrard, J. M. Goodman, B. Distel, H. Tabak and S. Subramani, Peroxisomal protein import is conserved between yeast, plants, insects and mammals., EMBO J. 9, 85–90 (1990).PubMedGoogle Scholar
  19. 19.
    P. J. Kraulis, MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures, J. Appl. Crystallogr. 24, 946–950 (1991).CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Peter Metcalf
    • 1
  • Martin Fusek
    • 2
  1. 1.EMBLHeidelbergGermany
  2. 2.Institute of Organic Chemistry and BiochemistryPragueCzech Republic

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