A camelid antibody fragment inhibits the formation of amyloid fibrils by human lysozyme (original) (raw)

• Letter
• Published: 14 August 2003
• Alexander M. Last2,
• Aline Desmyter3 nAff8,
• Klaas Decanniere3,
• Denis Canet2 nAff9,
• Göran Larsson1,
• Andrew Spencer4,
• David B. Archer5,
• Jurgen Sasse6,
• Serge Muyldermans3,
• Lode Wyns3,
• Christina Redfield2,
• André Matagne7,
• Carol V. Robinson1 &
• …
• Christopher M. Dobson1

Nature volume 424, pages 783–788 (2003)Cite this article

• 2697 Accesses
• 11 Altmetric
• Metrics details

Abstract

Amyloid diseases are characterized by an aberrant assembly of a specific protein or protein fragment into fibrils and plaques that are deposited in various organs and tissues1,2,3, often with serious pathological consequences. Non-neuropathic systemic amyloidosis4,5,6 is associated with single point mutations in the gene coding for human lysozyme. Here we report that a single-domain fragment of a camelid antibody7,8,9 raised against wild-type human lysozyme inhibits the in vitro aggregation of its amyloidogenic variant, D67H. Our structural studies reveal that the epitope includes neither the site of mutation nor most residues in the region of the protein structure that is destabilized by the mutation. Instead, the binding of the antibody fragment achieves its effect by restoring the structural cooperativity characteristic of the wild-type protein. This appears to occur at least in part through the transmission of long-range conformational effects to the interface between the two structural domains of the protein. Thus, reducing the ability of an amyloidogenic protein to form partly unfolded species can be an effective method of preventing its aggregation, suggesting approaches to the rational design of therapeutic agents directed against protein deposition diseases.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

• Purchase on SpringerLink
• Instant access to full article PDF

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

• Log in
• Learn about institutional subscriptions
• Read our FAQs
• Contact customer support

Similar content being viewed by others

References

1. Tan, S. Y. & Pepys, M. Amyloidosis. Histopathology 25, 403–414 (1994)
   Article CAS Google Scholar
2. Koo, E. H., Lansbury, P. T. Jr & Kelly, J. W. Amyloid diseases: Abnormal protein aggregation in neurodegeneration. Proc. Natl Acad. Sci. USA 96, 9989–9990 (1999)
   Article ADS CAS Google Scholar
3. Dobson, C. M. The structural basis of protein folding and its links with human disease. Phil. Trans. R. Soc. Lond. B 356, 133–145 (2001)
   Article CAS Google Scholar
4. Pepys, M. B. et al. Human lysozyme gene mutations cause hereditary systemic amyloidosis. Nature 362, 553–557 (1993)
   Article ADS CAS Google Scholar
5. Valleix, S. et al. Hereditary renal amyloidosis caused by a new variant lysozyme W64R in a French family. Kidney Int. 61, 907–912 (2002)
   Article CAS Google Scholar
6. Yazaki, M., Farrell, S. A. & Benson, M. D. A novel lysozyme mutation Phe57Ile associated with hereditary renal amyloidosis. Kidney Int. 63, 1652–1657 (2003)
   Article CAS Google Scholar
7. Dumoulin, M. et al. Single-domain antibody fragments with high conformational stability. Protein Sci. 11, 500–515 (2002)
   Article CAS Google Scholar
8. Hamers-Casterman, C. et al. Naturally occurring antibodies devoid of light chains. Nature 363, 446–448 (1993)
   Article ADS CAS Google Scholar
9. Muyldermans, S. Single domain camel antibodies: current status. J Biotechnol. 74, 277–302 (2001)
   CAS PubMed Google Scholar
10. Booth, D. R. et al. Instability, unfolding and aggregation of human lysozyme variants underlying amyloid fibrillogenesis. Nature 385, 787–793 (1997)
    Article ADS CAS Google Scholar
11. Canet, D. et al. Local cooperativity in the unfolding of an amyloidogenic variant of human lysozyme. Nature Struct. Biol. 9, 308–315 (2002)
    Article CAS Google Scholar
12. Schenk, D. Amyloid-beta immunotherapy for Alzheimer's disease: the end of the beginning. Nature Rev. Neurosci. 3, 824–828 (2002)
    Article CAS Google Scholar
13. Peretz, D. et al. Antibodies inhibit prion propagation and clear cell cultures of prion infectivity. Nature 412, 739–743 (2001)
    Article ADS CAS Google Scholar
14. White, A. R. et al. Monoclonal antibodies inhibit prion replication and delay the development of prion disease. Nature 422, 80–83 (2003)
    Article ADS CAS Google Scholar
15. Harper, J. D. & Lansbury, P. T. Jr Models of amyloid seeding in Alzheimer's disease and scrapie: mechanistic truths and physiological consequences of the time-dependent solubility of amyloid proteins. Annu. Rev. Biochem. 66, 385–407 (1997)
    Article CAS Google Scholar
16. Klunk, W. E., Pettegrew, J. W. & Abraham, D. J. Two simple methods for quantifying low-affinity dye-substrate binding. J. Histochem. Cytochem. 37, 1293–1297 (1989)
    Article CAS Google Scholar
17. Colon, W. & Kelly, J. W. Partial denaturation of transthyretin is sufficient for amyloid fibril formation in vitro. Biochemistry 31, 8654–8660 (1992)
    Article CAS Google Scholar
18. Morozova-Roche, L. A. et al. Amyloid fibril formation and seeding by wild-type human lysozyme and its disease-related mutational variants. J. Struct. Biol. 130, 339–351 (2000)
    Article CAS Google Scholar
19. Sunde, M. et al. Common core structure of amyloid fibrils by synchrotron X-ray diffraction. J. Mol. Biol. 273, 729–739 (1997)
    Article CAS Google Scholar
20. Aguzzi, A., Glatzel, M., Montrasio, F., Prinz, M. & Heppner, F. L. Interventional strategies against prion diseases. Nature Rev. Neurosci. 2, 745–749 (2001)
    Article CAS Google Scholar
21. Wolfe, M. S. Therapeutic strategies for Alzheimer's disease. Nature Rev. Drug Discov. 1, 859–866 (2002)
    Article CAS Google Scholar
22. Pepys, M. B. et al. Targeted pharmacological depletion of serum amyloid P component for treatment of human amyloidosis. Nature 417, 254–259 (2002)
    Article ADS CAS Google Scholar
23. Dobson, C. M. Protein folding and disease: a view from the first Horizon Symposium. Nature Rev. Drug Discov. 2, 154–160 (2003)
    Article CAS Google Scholar
24. May, B. C. et al. Potent inhibition of scrapie prion replication in cultured cells by bis-acridines. Proc. Natl Acad. Sci. USA 100, 3416–3421 (2003)
    Article ADS CAS Google Scholar
25. McCammon, M. G. et al. Screening transthyretin amyloid fibril inhibitors. Characterization of novel multiprotein, multiligand complexes by mass spectrometry. Structure (Cambridge) 10, 851–863 (2002)
    Article CAS Google Scholar
26. Hammarstrom, P., Wiseman, R. L., Powers, E. T. & Kelly, J. W. Prevention of transthyretin amyloid disease by changing protein misfolding energetics. Science 299, 713–716 (2003)
    Article ADS Google Scholar
27. Branden, C. & Tooze, J. Introduction to Protein Structure (Garland, New York, 1999)
    Google Scholar
28. Spencer, A. et al. Expression, purification, and characterization of the recombinant calcium-binding equine lysozyme secreted by the filamentous fungus Aspergillus niger: Comparisons with the production of hen and human lysozymes. Protein Expr. Purif. 16, 171–180 (1999)
    Article CAS Google Scholar
29. Ohkubo, T., Taniyama, Y. & Kikuchi, M. 1H and 15N NMR study of human lysozyme. J. Biochem. (Tokyo) 110, 1022–1029 (1991)
    Article CAS Google Scholar
30. Koradi, R., Billeter, M. & Wuthrich, K. MOLMOL: a program for display and analysis of macromolecular structures. J. Mol. Graph. 14, 51–55 (1996)
    Article CAS Google Scholar

Download references

Acknowledgements

The assistance of J. Zurdo, M. Krebs and B. Luisi for TEM and X-diffraction analysis of fibrils is gratefully acknowledged. We thank J.-M. Frère for many discussions. M.D. and D.C. were supported by a fellowship from the European Community. G.L. was supported by a fellowship from the Wenner-Gren Foundation. C.R. was supported by a BBSRC Advanced Research Fellowship. A.M. is a Research Associate of the FNRS, and was supported in part by a grant from the FRFC. C.V.R. is a Royal Society University Research Fellow. The research of C.M.D. is supported in part by a Programme Grant from the Wellcome Trust. This work was also supported by a BBSRC grant (to C.M.D., C.V.R. and D.B.A.) and by the Belgian Government through the PAI.

Author information

Author notes

1. Aline Desmyter
   Present address: Centre National de la Recherche Scientifique, 31 Chemin Joseph Aiguier, 13402, Marseille, France
2. Denis Canet
   Present address: Gene Prot Inc., 2 Pré-de-la-Fontaine, 1217, Meyrin/GE, Switzerland

Authors and Affiliations

1. Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW, Cambridge, UK
   Mireille Dumoulin, Göran Larsson, Carol V. Robinson & Christopher M. Dobson
2. Oxford Centre for Molecular Sciences, University of Oxford, South Parks Road, OX1 3QH, Oxford, UK
   Alexander M. Last, Denis Canet & Christina Redfield
3. Department Ultrastructure, Vrije Universiteit Brussel, Pleinlaan 2, B-1050, Brussels, Belgium
   Aline Desmyter, Klaas Decanniere, Serge Muyldermans & Lode Wyns
4. Institute of Food Research, Norwich Research Park, Colney, NR4 7UA, Norwich, UK
   Andrew Spencer
5. School of Life and Environmental Sciences, University of Nottingham, University Park, NG7 2RD, Nottingham, UK
   David B. Archer
6. Central Veterinary Research Laboratory, P.O. Box 597, Dubai, United Arab Emirates
   Jurgen Sasse
7. Laboratoire d'Enzymologie, Centre d'Ingénerie des Protéines, Institut de Chimie B6, Université de Liège, B-4000, Sart Tilman, Liège, Belgium
   André Matagne

Authors

1. Mireille Dumoulin
   You can also search for this author inPubMed Google Scholar
2. Alexander M. Last
   You can also search for this author inPubMed Google Scholar
3. Aline Desmyter
   You can also search for this author inPubMed Google Scholar
4. Klaas Decanniere
   You can also search for this author inPubMed Google Scholar
5. Denis Canet
   You can also search for this author inPubMed Google Scholar
6. Andrew Spencer
   You can also search for this author inPubMed Google Scholar
7. David B. Archer
   You can also search for this author inPubMed Google Scholar
8. Jurgen Sasse
   You can also search for this author inPubMed Google Scholar
9. Serge Muyldermans
   You can also search for this author inPubMed Google Scholar
10. Lode Wyns
    You can also search for this author inPubMed Google Scholar
11. Christina Redfield
    You can also search for this author inPubMed Google Scholar
12. André Matagne
    You can also search for this author inPubMed Google Scholar
13. Carol V. Robinson
    You can also search for this author inPubMed Google Scholar
14. Christopher M. Dobson
    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence toChristopher M. Dobson.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Rights and permissions

About this article

Cite this article

Dumoulin, M., Last, A., Desmyter, A. et al. A camelid antibody fragment inhibits the formation of amyloid fibrils by human lysozyme.Nature 424, 783–788 (2003). https://doi.org/10.1038/nature01870

Download citation

• Received: 22 March 2003
• Accepted: 18 June 2003
• Issue Date: 14 August 2003
• DOI: https://doi.org/10.1038/nature01870