5E0K: X-ray Crystal Structure Of Tryptophan Synthase Complex From Pyrococcus Furiosus At 2.76 A

Enzymes in heteromeric, allosterically regulated complexes catalyze a rich array of chemical reactions. Separating the subunits of such complexes, however, often severely attenuates their catalytic activities, because they can no longer be activated by their protein partners. We used directed evolution to explore allosteric regulation as a source of latent catalytic potential using the beta-subunit of tryptophan synthase from Pyrococcus furiosus (PfTrpB). As part of its native alphabetabetaalpha complex, TrpB efficiently produces tryptophan and tryptophan analogs; activity drops considerably when it is used as a stand-alone catalyst without the alpha-subunit. Kinetic, spectroscopic, and X-ray crystallographic data show that this lost activity can be recovered by mutations that reproduce the effects of complexation with the alpha-subunit. The engineered PfTrpB is a powerful platform for production of Trp analogs and for further directed evolution to expand substrate and reaction scope.
PDB ID: 5E0KDownload
MMDB ID: 134156
PDB Deposition Date: 2015/9/29
Updated in MMDB: 2015/11
Experimental Method:
x-ray diffraction
Resolution: 2.76  Å
Source Organism:
Similar Structures:
Biological Unit for 5E0K: tetrameric; determined by author and by software (PISA)
Molecular Components in 5E0K
Label Count Molecule
Proteins (4 molecules)
Tryptophan Synthase Alpha Chain(Gene symbol: PF_RS08610)
Molecule annotation
Tryptophan Synthase Beta Chain 1(Gene symbol: PF_RS08615)
Molecule annotation
Chemicals (2 molecules)
* Click molecule labels to explore molecular sequence information.

Citing MMDB