1d6f
From Proteopedia
| Line 1: | Line 1: | ||
[[Image:1d6f.jpg|left|200px]] | [[Image:1d6f.jpg|left|200px]] | ||
| - | + | <!-- | |
| - | + | The line below this paragraph, containing "STRUCTURE_1d6f", creates the "Structure Box" on the page. | |
| - | + | You may change the PDB parameter (which sets the PDB file loaded into the applet) | |
| - | + | or the SCENE parameter (which sets the initial scene displayed when the page is loaded), | |
| - | + | or leave the SCENE parameter empty for the default display. | |
| - | | | + | --> |
| - | | | + | {{STRUCTURE_1d6f| PDB=1d6f | SCENE= }} |
| - | + | ||
| - | + | ||
| - | }} | + | |
'''CHALCONE SYNTHASE C164A MUTANT''' | '''CHALCONE SYNTHASE C164A MUTANT''' | ||
| Line 31: | Line 28: | ||
[[Category: Jez, J M.]] | [[Category: Jez, J M.]] | ||
[[Category: Noel, J P.]] | [[Category: Noel, J P.]] | ||
| - | [[Category: | + | [[Category: Flavonoid biosynthesis]] |
| - | [[Category: | + | [[Category: Malonyl-coa decarboxylation]] |
| - | [[Category: | + | [[Category: Polypetide synthase]] |
| - | [[Category: | + | [[Category: Site- directed mutant]] |
| - | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 13:30:04 2008'' | |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | |
Revision as of 10:30, 2 May 2008
CHALCONE SYNTHASE C164A MUTANT
Overview
Chalcone synthase (CHS) catalyzes formation of the phenylpropanoid chalcone from one p-coumaroyl-CoA and three malonyl-coenzyme A (CoA) thioesters. The three-dimensional structure of CHS [Ferrer, J.-L., Jez, J. M., Bowman, M. E., Dixon, R. A., and Noel, J. P. (1999) Nat. Struct. Biol. 6, 775-784] suggests that four residues (Cys164, Phe215, His303, and Asn336) participate in the multiple decarboxylation and condensation reactions catalyzed by this enzyme. Here, we functionally characterize 16 point mutants of these residues for chalcone production, malonyl-CoA decarboxylation, and the ability to bind CoA and acetyl-CoA. Our results confirm Cys164's role as the active-site nucleophile in polyketide formation and elucidate the importance of His303 and Asn336 in the malonyl-CoA decarboxylation reaction. We suggest that Phe215 may help orient substrates at the active site during elongation of the polyketide intermediate. To better understand the structure-function relationships in some of these mutants, we also determined the crystal structures of the CHS C164A, H303Q, and N336A mutants refined to 1.69, 2.0, and 2.15 A resolution, respectively. The structure of the C164A mutant reveals that the proposed oxyanion hole formed by His303 and Asn336 remains undisturbed, allowing this mutant to catalyze malonyl-CoA decarboxylation without chalcone formation. The structures of the H303Q and N336A mutants support the importance of His303 and Asn336 in polarizing the thioester carbonyl of malonyl-CoA during the decarboxylation reaction. In addition, both of these residues may also participate in stabilizing the tetrahedral transition state during polyketide elongation. Conservation of the catalytic functions of the active-site residues may occur across a wide variety of condensing enzymes, including other polyketide and fatty acid synthases.
About this Structure
1D6F is a Single protein structure of sequence from Medicago sativa. Full crystallographic information is available from OCA.
Reference
Dissection of malonyl-coenzyme A decarboxylation from polyketide formation in the reaction mechanism of a plant polyketide synthase., Jez JM, Ferrer JL, Bowman ME, Dixon RA, Noel JP, Biochemistry. 2000 Feb 8;39(5):890-902. PMID:10653632 Page seeded by OCA on Fri May 2 13:30:04 2008
