Site-directed mutagenesis of the active site serine290 in flavanone 3β- hydroxylase from Petunia hybrida

European Journal of Biochemistry

Volumn 267, Issue 3, 2000, Pages 853-860

  Lukačin, Richard ^a   Gröning, Inga ^a   Pieper, Uwe ^b   Matern, Ulrich ^a  

^a PHILIPPS UNIVERSITY MARBURG   (Germany)

^b JUSTUS LIEBIG UNIVERSITY   (Germany)

Author keywords

2 oxoglutarate binding site; Flavanone 3 hydroxylase; Flavonoid biosynthesis; Petunia hybrida; Site directed mutagenesis

Indexed keywords

2 OXOGLUTARIC ACID; ALANINE; AMINO ACID; CARBON DIOXIDE; CATECHIN; DEACETOXYCEPHALOSPORIN C; ENZYME; FLAVANONE; FLAVONOID; FLAVONOL DERIVATIVE; ISOPENICILLIN N; OXYGENASE; PROANTHOCYANIDIN; SERINE; SUCCINIC ACID; SYNTHETASE; THREONINE; VALINE;

ARTICLE; BINDING SITE; BIOSYNTHESIS; CATALYSIS; CIRCULAR DICHROISM; DECARBOXYLATION; ENZYME ACTIVE SITE; NONHUMAN; PLANT; PRIORITY JOURNAL; SITE DIRECTED MUTAGENESIS;

AMINO ACID SEQUENCE; BASE SEQUENCE; CATALYTIC DOMAIN; CIRCULAR DICHROISM; CONSERVED SEQUENCE; DNA PRIMERS; ESCHERICHIA COLI; KINETICS; MIXED FUNCTION OXYGENASES; MOLECULAR SEQUENCE DATA; MUTAGENESIS, SITE-DIRECTED; PROTEIN CONFORMATION; RECOMBINANT PROTEINS; SEQUENCE HOMOLOGY, AMINO ACID; SERINE; SOLANACEAE;

ESCHERICHIA COLI; PETUNIA; PETUNIA X HYBRIDA;

EID: 0038632837     PISSN: 00142956     EISSN: None     Source Type: Journal    
DOI: 10.1046/j.1432-1327.2000.01064.x     Document Type: Article

References (38)

1. 1. Heller, W. & Forkmann, G. (1993) Biosynthesis of flavonoids. In The Flavonoids, Advances in Research Since 1986 (Harborne, J.B., eds), pp. 399-425. Chapman & Hall, London.
2. 2. Prescott, A.G.J. (1993) A dilemma of dioxygenases (or where biochemistry and molecular biology fail to meet). J. Exp. Bot. 44, 849-861.
3. 3. De Carolis, E. & Luca, V. (1994) 2-Oxoglutarate-dependent dioxygenase and related enzymes: biochemical characterization. Phytochemistry 36, 1093-1107.
4. 4. Britsch, L., Ruhnau-Brich, B. & Forkmann, G. (1992) Molecular cloning, sequence analysis, and in vitro expression of flavanone 3β-hydroxylase from Petunia hybrida. J. Biol. Chem. 267, 5380-5387.
5. 5. Britsch, L., Dedio, J., Saedler, H. & Forkmann, G. (1993) Molecular characterization of flavanone 3β-hydroxylase: consensus sequence, comparison with related enzymes and the role of conserved histidines. Eur. J. Biochem. 217, 745-754.
6. 6. Holton, T.A., Brugliera, F. & Tanaky, Y. (1993) Cloning and expression of flavonol synthase from Petunia hybrida. Plant J. 4, 1003-1010.
7. 7. Suzuki, K., Yun, D.J., Chen, X.Y., Yamada, Y. & Hashimoto, T. (1999) An Atropa belladonna hyoscyamine 6β-hydroxylase gene is differentially expressed in the root pericycle and anthers. Plant. Mol. Biol. 40, 141-152.
8. 8. Roach, P.L., Clifton, I.J., Fülöp, V., Harlos, K., Barton, G.J., Hajdu, J., Andersson, I., Schofield, C.J. & Baldwin, J.E. (1995) Crystal structure of isopenicillin N-synthase is the first from a new structural family of enzymes. Nature 375, 700-704.
9. 9. Valegård, K., Van Schelting, T., Lloyd, M.D., Hara, T., Ramaswamy, S., Perrakis, A., Thompson, A., Lee, H.-J., Baldwin, J.E., Schofield, C.J., Hajdu, J. & Andersson, I. (1998) Structure of a cephalosporin synthase. Nature 394, 805-809.
10. 10. Roach, P.L., Clifton, I.J., Hensgens, C.M.H., Shibata, N., Schofield, C.J., Hajdu, J. & Baldwin, J.E. (1997) Structure of isopenicillin N synthase complexed with substrate and the mechanism of penicillin formation. Nature 387, 827-830.
11. 11. Lukačin, R. & Britsch, L. (1997) Identification of strictly conserved histidine and arginine residues as part of the active site in Petunia hybrida flavanone 3β-hydroxylase. Eur. J. Biochem. 249, 748-757.
12. 12. Loke, P. & Sim, T.-S. (1998) Mutational evidence for the role of serine-283 in Cephalosporium acremonium isopenicillin N synthase. FEMS Microbiol. Lett. 165, 353-356.
13. 13. Sanger, F., Nicklen, S. & Coulsen, A.R. (1977) DNA sequencing with chain terminating inhibitors. Proc. Natl Acad. Sci. USA 74, 5463-5467.
14. 14. Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
15. 15. Lukačin, R., Springob, K., Urbanke, C., Ernwein, C., Schröder, G., Schröder, J. & Matern, U. (1999) Native acridone synthases I and II from Ruta graveolens L. form homodimers. FEBS Lett. 448, 135-140.
16. 16. Lukačin, R., Gröning, I., Schiltz, E., Britsch, L. & Matern, U. (2000) Purification of recombinant flavanone 3β-hydroxylase from Petunia hybrida and assignment of primary site of proteolytic degradation. Arch. Biochem. Biophys. In press.
17. 17. Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 221, 680-685.
18. 18. Towbin, H., Staehelin, T. & Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl Acad. Sci. USA 76, 4350-4354.
19. 19. Britsch, L. (1990) Purification of flavanone 3β-hydroxylase from Petunia hybrida: antibody preparation and characterization of a chemogenetically defined mutant. Arch. Biochem. Biophys. 276, 348-354.
20. 20. Sreerama, N. & Woody, R.W. (1993) A self-consistent method for the analysis of protein secondary structure from circular dichroism. Anal. Biochem. 209, 32-44.
21. 21. Deleage, G. & Geourjon, C. (1993) An interactive graphic program for calculating the secondary structure content of proteins from circular dichroism spectrum. Comput. Appl. Biosci. 9, 197-199.
22. 55-isoprenoid alcohol phosphokinase from Staphylococcus aureus. J. Biol. Chem. 247, 5123-5131.
23. 23. Bensadoun, A. & Weinstein, B. (1976) Assay of proteins in the presence of interfering material. Anal. Biochem. 70, 241-250.
24. 24. Britsch, L. & Grisebach, H. (1986) Purification and characterization of (2S)-flavanone 3-hydroxylase from Petunia hybrida. Eur. J. Biochem. 156, 569-577.
25. 25. Huang, Y. & Dennis, D.T. (1995) Histidine residues 139, 363 and 500 are essential for catalytic activity of cofactor-independent phosphoglyceromutase from developing endosperm of the castor plant. Eur. J. Biochem. 229, 395-402.
26. 26. Galla, H.-J., Ed. (1988) Spektroskopische Methoden in der Biochemie, pp. 23-30. Georg Thieme-Verlag, Stuttgart.
27. 27. Roberge, M., Dupont, C., Morosoli, R., Shareck, F. & Kluepfel, D. (1997) Asparagine-127 of xylanase A from Streptomyces lividans, a key residue in glycosyl hydrolases of superfamily 4/7: kinetic evidence for its involvement in stabilization of the catalytic intermediate. Protein Eng. 10, 399-403.
28. 28. Borovok, I., Landman, O., Kreisberg-Zakarin, R., Aharonowitz, Y. & Cohen, G. (1996) Ferrous active site of isopenicillin N synthase: genetic and sequence analysis of the endogenous ligands. Biochemistry 35, 1981-1987.
29. 29. Durairaj, M., Leskiw, B.K. & Jensen, S.E. (1996) Genetic and biochemical analysis of the cysteinyl residues of isopenicillin N synthase from Streptomyces clavuligerus. Can. J. Microbiol. 42, 870-875.
30. 30. Baldwin, J.E. & Abraham, E.P. (1988) The biosynthesis of penicillins and cephalosporins. Nat. Prod. Rep. 5, 129-145.
31. 31. Forkmann, G., Heller, W. & Grisebach, H. (1980) Anthocyanin biosynthesis in flowers of Matthiola incana. Flavanone 3-and flavonoid 3′-hydroxylases. Z. Naturforsch., Sect. C Biosci. 35, 691-695.
32. 32. Britsch, L., Heller, W. & Grisebach, H. (1981) Conversion of flavanone to flavone, dihydroflavonol and flavonol with an enzyme system from cell cultures of parsley. Z. Naturforsch. Sect. C Biosci. 36, 742-750.
33. 33. Anzellotti, D. & Ibrahim, R.K. (1999) A novel α-ketoglutarate-dependent dioxygenase catalyzes the 6-hydroxylation of partially methylated flavonols in Chrysoplenium americanum (Poster). Proceedings of the Phytochemical Society of North America, Montreal, Canada, pp. 34-35.
34. 34. Hashimoto, T. & Yamada, Y. (1986) Hyoscyamine 6β-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, in alkaloid-producing root cultures. Plant Physiol. 81, 619-625.
35. 35. De Carolis, E., Chan, F., De Balsevich, J. & Luca, V. (1990) Isolation and characterization of a 2-oxoglutarate-dependent dioxygenase involved in the second-to-last step in vindoline biosynthesis. Plant Physiol. 94, 1323-1329.
36. 20-gibberellins. J. Plant Growth Regul. 1, 105-116.
37. 37. White, R.L., John, E.-M., Baldwin, J.E. & Abraham, E.P. (1982) Stoichiometry of oxygen consumption in the biosynthesis of isopenicillin from a tripeptide. Biochem. J. 203, 791-793.
38. 38. Dong, J.G., Fernandez-Maculet, J.C. & Yang, S.-F. (1992) Purification and characterization of 1-aminocyclopropane-1-carboxylate oxidase from apple fruit. Proc. Natl Acad. Sci. USA 89, 9789-9793.