Pentalenene synthase. Histidine-309 is not required for catalytic activity

Journal of the American Chemical Society

Volumn 121, Issue 3, 1999, Pages 591-592

  Seemann, Myriam ^a   Zhai, Guangzhi ^a   Umezawa, Kazuo ^a,b   Cane, David ^a  

^a BROWN UNIVERSITY   (United States)

^b KEIO UNIVERSITY   (Japan)

Author keywords

[No Author keywords available]

Indexed keywords

ALANINE; CYSTEINE; HISTIDINE; MUTANT PROTEIN; PENTALENENE; PENTALENENE SYNTHASE; PHENYLALANINE; SERINE; SESQUITERPENE; SYNTHETASE; UNCLASSIFIED DRUG;

AMINO ACID SEQUENCE; ARTICLE; ENZYME ACTIVE SITE; ENZYME KINETICS; NONHUMAN; NUCLEOTIDE SEQUENCE; SITE DIRECTED MUTAGENESIS;

EID: 0033608083     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja983657h     Document Type: Article

References (27)

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21. Germacrene A was a gift from Dr. Jan-Willem de Kraker (Wageningen Agricultural University, The Netherlands).
22. 3d the stereochemical course of the reactions catalyzed by the various histidine mutants has not yet been determined.
23. Chappell and Noel have suggested that an active-site tryptophan (W273) can serve as the base in the mechanistically closely related cyclization catalyzed by epi-aristolochene synthase, a protein closely related in three-dimensional structure but with no similarity in sequence to pentalenene synthase. Indeed, these researchers have found that replacement of W273 abolishes cyclase activity in epi-aristolochene synthase. (Starks, C. M.; Back, K.; Chappell, J.; Noel, J. P. Science 1997, 277, 1815-1820.) Interestingly the latter cyclization is believed to involve germacrene A (7) as a mandatory intermediate. By contrast, we have observed that the corresponding PS double mutant, H309F/W308F, retains substantial cyclase activity (M. Seemann, unpublished observations.)
24. Chappell and Noel have suggested that an active-site tryptophan (W273) can serve as the base in the mechanistically closely related cyclization catalyzed by epi-aristolochene synthase, a protein closely related in three-dimensional structure but with no similarity in sequence to pentalenene synthase. Indeed, these researchers have found that replacement of W273 abolishes cyclase activity in epi-aristolochene synthase. (Starks, C. M.; Back, K.; Chappell, J.; Noel, J. P. Science 1997, 277, 1815-1820.) Interestingly the latter cyclization is believed to involve germacrene A (7) as a mandatory intermediate. By contrast, we have observed that the corresponding PS double mutant, H309F/W308F, retains substantial cyclase activity (M. Seemann, unpublished observations.)
25. We have previously reported the formation of anomalous cyclization products by mutants of trichodiene synthase: Cane. D. E.; Xue, Q. J. Am. Chem. Soc. 1996, 118, 1563-1564. Cane, D. E.; Xue. Q.; Fitzsimons, B. C. Biochemistry 1996, 35, 12369-12376. Cane, D. E.; Xue, Q.; Van Epp, J. E. J. Am. Chem. Soc. 1996, 118, 8499-8500.
26. We have previously reported the formation of anomalous cyclization products by mutants of trichodiene synthase: Cane. D. E.; Xue, Q. J. Am. Chem. Soc. 1996, 118, 1563-1564. Cane, D. E.; Xue. Q.; Fitzsimons, B. C. Biochemistry 1996, 35, 12369-12376. Cane, D. E.; Xue, Q.; Van Epp, J. E. J. Am. Chem. Soc. 1996, 118, 8499-8500.
27. We have previously reported the formation of anomalous cyclization products by mutants of trichodiene synthase: Cane. D. E.; Xue, Q. J. Am. Chem. Soc. 1996, 118, 1563-1564. Cane, D. E.; Xue. Q.; Fitzsimons, B. C. Biochemistry 1996, 35, 12369-12376. Cane, D. E.; Xue, Q.; Van Epp, J. E. J. Am. Chem. Soc. 1996, 118, 8499-8500.