Evolution of enzymatic activities in the enolase superfamily: Identification of a 'new' general acid catalyst in the active site of D- galactonate dehydratase from Escherichia coli

Journal of the American Chemical Society

Volumn 121, Issue 18, 1999, Pages 4540-4541

  Wieczorek, Stacey J ^b   Kalivoda, Kathryn A ^b   Clifton, James G ^a   Ringe, Dagmar ^a   Petsko, Gregory A ^a   Gerlt, John A ^b  

^a BRANDEIS UNIVERSITY   (United States)

^b NONE

Author keywords

[No Author keywords available]

Indexed keywords

BACTERIAL ENZYME; DEXTRO GALACTONATE DEHYDRATASE; ENOLASE; HYDROLYASE; UNCLASSIFIED DRUG;

ARTICLE; CATALYST; ENZYME ACTIVE SITE; ENZYME MECHANISM; ENZYME SUBSTRATE; ESCHERICHIA COLI; NONHUMAN; PROTEIN FAMILY; SEQUENCE ANALYSIS; STEREOSPECIFICITY;

EID: 0033549133     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja990500w     Document Type: Article

References (14)

1. Babbitt, P. C.; Hasson, M.; Wedekind, J. E.; Palmer, D. R. J.; Rayment, I.; Ringe, D.; Kenyon, G. L.; Gerlt, J. A. Biochemistry 1996, 35, 16489-16501.
2. Palmer, D. R. J.; Gerlt, J. A. J. Am. Chem. Soc. 1996, 118, 10323-10324.
3. Palmer, D. R. J.; Hubbard, B. K.; Gerlt, J. A. Biochemistry 1998, 37, 14350-14357.
4. Hubbard, B. K.; Koch, M.; Palmer, D. R. J.; Gerlt, J. A. Biochemistry 1998, 37, 143690-14375.
5. Hubbard, B. K.; Delli, J.; Gerlt, J. A., unpublished observations.
6. Babbitt, P. C.; Mrachko, G. T.; Hasson, M. S.; Huisman, G. W.; Kolter, R.; Ringe, D.; Petsko, G. A.; Kenyon, G. L.; Gerlt, J. A. Science 1995, 267, 1159-1161.
7. Gulick, A. M.; Palmer, D. R. J.; Babbitt, P. C.; Gerlt, J. A.; Rayment, I. Biochemistry 1998, 37, 14358-14368.
8. The conversion of D-galactonate (1) and 3-FGal (6) to 2-keto-3-deoxy-D-galactonate was quantitated using a semicarbazide assay (MacGee, J.; Doudoroff, M. J. Biol. Chem. 1954, 210, 617-624).
9. Clifton, J. G.; Wieczorek, S. J.; Gerlt, J. A.; Petsko, G. A., experiments in progress. The current structure with eight polypeptides in the asymmetric unit is refined at 3.2 Å resolution. The effective resolution of the structure was increased by 8-fold averaging of the electron density map. A full description of the structure will be published and coordinates deposited in the Protein Data Bank when refinement at higher resolution is completed. Until that time, coordinates are available without restriction by request to G.A.P. (E-mail: petsko@binah.cc.brandeis.edu).
10. 2O) δ-207 (m).
11. Palmer, D. R. J.; Wieczorek, S. J.; Hubbard, B. K.; Mrachko, G. T.; Gerlt, J. A. J. Am. Chem. Soc. 1997, 119, 9580-9581.
12. Babbitt, P. C.; Gerlt, J. A. J. Biol. Chem. 1997, 272, 30591-30594.
13. RspA from E. coli, SwissProt P38104; Yin2 from Streptomyces ambofaciens, SwissProt P32436; and SpaA from Streptomyces coelicolor, PIR JC5178. RspA, Yin2, and SpaA each share ∼32% sequence identity with Ga1D; the sequence identities relating RspA, Yin2, and SpaA are ≥60%, suggesting that these proteins may catalyze the same reaction.
14. Supported by NIH GM-40570 (to J.A.G., G.A.P.) and GM-52594 (to J.A.G.). We thank Dr. Patricia C. Babbitt, University of California, San Francisco, for helpful discussions.