Antibody-catalyzed retro-aldol reaction

Journal of the American Chemical Society

Volumn 118, Issue 25, 1996, Pages 6078-6079

  Flanagan, Mark E ^a   Jacobsen, John R ^a   Sweet, Elizabeth ^a   Schultz, Peter G ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ANTIBODY;

ARTICLE; DRUG MANUFACTURE; ENZYME ACTIVITY; MODEL; REACTION ANALYSIS;

EID: 0030056177     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja954221u     Document Type: Article

References (31)

1. Marsh, J. J.; Lebherz, H. G. Trends Biochem. Sci. 1992, 17, 110.
2. (a) Heathcock, C. H. Aldrichimica Acta 1990, 23, 99.
3. (b) Evans, D. A. Science 1988, 240, 420.
4. (a) Tramontano, A.; Janda, K. D.; Lerner, R. A. Science 1986, 234, 1566.
5. (b) Pollack, S. J.; Jacobs, J. W.; Schultz, P. G. Science 1986, 234, 1570.
6. (a) Schultz, P. G.; Lerner, R. A. Acc. Chem. Res. 1993, 26, 391.
7. (b) Schultz, P. G.; Lerner R. A. Science 1995, 269, 1835.
8. (a) Reymond, J.-L.; Chen, Y. Tetrahedron Lett. 1995, 36, 2575.
9. (b) Koch, T.; Reymond, J.-L.; Lerner, R. A. J. Am. Chem. Soc. 1995, 117, 9383.
10. (c) Wagner, J.; Lerner, R. A.; Barbas, C. F., III Science 1995, 270, 1797.
11. Henry, L. C. R. Acad. Sci. Paris 1895, 120, 1265.
12. (a) Jencks, W. P. Catalysis in Chemistry and Enzymology; Dover: Mineola, NY, 1987; p 11501.
13. (b) Fersht, A. Enzyme Structure and Mechanism; W. H. Freeman: New York, NY, 1985.
14. 370 = 22 900 for the azo linkage and determined to be 14 for KLH and 8 for BSA.
15. (a) Harlow, E.; Lane, D. Antibodies: A Laboratory Manual; Cold Spring Harbor Laboratory: USA, 1988.
16. (b) Shokat, K. M. Ph.D. Thesis, University of California, Berkeley, CA, 1991.
17. Fredriksson, U.-B.; Fagerstam, L. G.; Cole, A. W. G.; Lundgren, T. Protein A-Sepharose C1-4B Affinity Purification of IgG Monoclonal Antibodies from Mouse Ascites; Pharmacia AB: Uppsala, Sweden, 1986.
18. Antibodies were assayed for catalysis of the aldol condensation using reaction mixtures of benzaldehyde and phenylacetone at pH 8.0 and 9.0. 4-Fluorophenylacetone as well as the ketone containing an ortho azo linkage to p-cresol was assayed in pH 9.0 buffer.
19. A derivative of hapten 10 was generated by diazotization and coupling to p-cresol. This analog was expected to have greater structural homology to the hapten-protein conjugate used for immunization and screening. No inhibition of catalysis was observed at concentrations up to 500 μM.
20. (a) Wittig, G.; Reiff, H. Angew. Chem., Int. Ed. Engl. 1968, 7, 7.
21. (b) Wittig, G.; Suchanek, P. Tetrahedron Suppl. 1967, 8, 347.
22. Alberty, W. J. Adv. Phys. Org. Chem. 1993, 28, 139.
23. f = 0.25).
24. The relative stereochemical configuration of syn-4 was determined by single-crystal X-ray analysis.
25. No evidence of enantioselectivity was observed for catalysis by monitoring the relative consumption of the enantiomers of syn-4 by chiral HPLC (Chiralcel OD-R column, 30-70% acetonitrile in 50 mM NaOAc, pH 5.0).
26. (a) Cha, S. Biochem. Pharmacol. 1975, 24, 2177.
27. (b) Angeles, T. S. et al. Biochemistry 1993, 32, 12128.
28. Reactions were carried out in 50 mM NaOAc, 50 mM NaCl, 5% acetonitrile, pH 5.0 with 50 μM syn-4 and 3 μM 29C5.1, 4 °C.
29. Holbrook, J. J.; Ingram, V. A. Biochem. 1973, 131, 729.
30. -1 at pH 5.0).
31. Enzyme digests for Fab preparation used immobilized Papain (Pierce). Isooctane reactions were conducted in the presence of 1.0% Aerosol OT (Sigma) as described by Paradkar, V. M.; Dordick, J. S. J. Am. Chem. Soc. 1994, 116, 5009.