Abstract
BIOLOGICAL macromolecules with catalytic activity can be created artificially using two approaches. The first exploits a system that selects a few catalytically active biomolecules from a large pool of randomly generated (and largely inactive) molecules. Catalytic antibodies1 and many catalytic RNA molecules2 are obtained in this way. The second involves rational design of a biomolecule that folds in solution to present to the substrate an array of catalytic functional groups3–8. Here we report the synthesis of rationally designed polypeptides that catalyse the decarboxylation of oxaloacetate via an imine intermediate. We determine the secondary structures of the polypeptides by two-dimensional NMR spectroscopy. We are able to trap and identify intermediates in the catalytic cycle, and to explore the kinetics in detail. The formation of the imine by our artificial oxaloacetate decarboxylases is three to four orders of magnitude faster than can be achieved with simple amine catalysts: this performance rivals that of typical catalytic antibodies.
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 Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Lerner, R. A., Benkovic, S. J. & Schultz, P. G. Science 252, 659–667 (1991).
Bartel, D. P. & Szostak, J. W. Science 261, 1411–1416 (1993).
Sheehan, J. C., Bennett, G. B. & Schneider, J. A. J. Am. chem. Soc. 88, 3455–3456 (1966).
Chakravarty, P. K., Mathur, K. B. & Dhar, M. M. Experientia 29, 786–788 1973).
Gutte, B., Daeumigen, M. & Wittschieber, E. Nature 281, 650–655 (1979).
Langen, H. et al. Eur. J. Biochem. 182, 727–735 (1989).
Sasaki, T. & Kaiser, E. T. J. Am. chem. Soc 111, 380–381 (1989).
Hahn, K. W., Klis, W. A. & Stewart, J. Science 248, 1544–1546 (1990).
Rozzell, J. D. Jr Meth. Enzym. 136, 479–503 (1987).
Seltzer, S., Hamilton, G. A. & Westheimer, F. H. J. Am. chem. Soc. 81, 4018–4024 (1959).
Hay, R. W. Aust, J. Chem. 18, 337–351 (1965).
Guthrie, J. P. & Jordan, F. J. Am. chem. Soc. 94, 9132–9136 (1972); J. Am. chem. Soc. 94, 9136–9141 (1972).
Leussing, D. L. & Raghavan, N. V. J. Am. chem. Soc. 102, 5635–5643 (1980).
Westheimer, F. H. Proc. Robert Welch Fdn Conf. chem. Res. Vol. 15 7–50 (Robert A. Welch Foundation, Houston, 1971).
Rozzell, J. D. Jr & Benner, S. A. J. Am. chem. Soc. 106, 4937–4941 (1984).
Benner, S. A., Glasfeld, A. & Piccirilli, J. A. Top. Stereochem 127–207 (1989).
Jencks, W. P. Catalysis in Chemistry and Enzymology 587 (McGraw-Hill, New York, 1969).
Johnsson, K. thesis, Eidgenössische Technische Hochschule (1992).
Koehler, K., Sandstrom, W. & Cordes, E. H. J. Am. chem. Soc. 86, 2413–2419 (1964).
Reimann, J. P. & Jencks, W. P. J. Am. chem. Soc. 88, 3973–3982 (1966).
Jencks, W. P. Adv. Enzym. 43, 219–410 (1975).
Hine, J. & Via, F. H. J. Am. chem. Soc. 94, 190–194 (1972).
Kaiser, E. T. in Redesigning the Molecules of Life (eds Benner, S. A.) 25–28 (Springer, Heidelberg, 1988).
Eisenberg, D. et al. Proteins 1, 16–22 (1986).
Ho, S. P. & DeGrado, W. F. J. Am. chem. Soc. 109, 6751–6758 (1987).
Goraj, K., Renard, A., Martial, J. A. Protein Engng 3, 259–266 (1990).
Ghadiri, M. R., Scares, C. & Choi, C. J. Am. chem. Soc. 114, 825–831 (1992).
Hill, C. P., Anderson, D. A., Wesson, L., DeGrado, W. F. & Eisenberg, D. Science 249, 543–546 (1990).
Allemann, R. K. thesis, Eidgenössische Technische Hochschule (1989).
Wada, A. Adv. Biophys. 9, 1–63 (1976).
Hoi, W. G. J., van Duijnen, P. T. & Berendsen, H. J. C. Nature 273, 443–446 (1978).
Sali, D., Bycroft, M. & Fersht, A. R. Nature 335, 740–743 (1988).
Wüthrich, K. NMR of Proteins and Nucleic Acids (Wiley, New York, 1986).
Nelson, J. W. & Kallenbach, N. P. Proteins 1, 211–217 (1986).
O'Leary, M. & Westheimer, F. H. Biochemistry, 7, 913–919 (1968).
Kaplan, H., Stephenson, K. J. & Hartley, B. S. Biochem. J. 124, 289–299 (1971).
Pollack, S. J., Jacobs, J. W. & Schultz, P. G. Science 234, 1570–1573 (1986).
Spetnagel, W. J. & Klotz, I. M. J. Am. chem. Soc. 98, 8199–8204 (1976).
Bruice, P. H. J. Am. chem. Soc. 111, 962–970 (1989).
Merrifield, R. B. & Barany, G. in The Peptides, Analysis, Synthesis, Biology Vol. 2 (eds Gross, E. & Meienhofer, J.) 3–284 (Academic, New York, 1980).
Johnsson, K., Allemann, R. K. & Benner, S. A. in Molecular Mechanisms in Bioorganic Processes (eds Bleasedale, C. & Golding, B. T.) 166–167 (Royal Society of Chemistry, Cambridge, 1990).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Johnsson, K., Allemann, R., Widmer, H. et al. Synthesis, structure and activity of artificial, rationally designed catalytic polypeptides. Nature 365, 530–532 (1993). https://doi.org/10.1038/365530a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/365530a0
This article is cited by
-
A photochemical dehydrogenative strategy for aniline synthesis
Nature (2020)
-
Catalytic diversity in self-propagating peptide assemblies
Nature Chemistry (2017)
-
Simple peptides derived from the ribosomal core potentiate RNA polymerase ribozyme function
Nature Chemistry (2017)
-
Effect of side chain length on intrahelical interactions between carboxylate- and guanidinium-containing amino acids
Amino Acids (2014)
-
Designing artificial enzymes by intuition and computation
Nature Chemistry (2010)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.
