5E0K

X-ray crystal structure of tryptophan synthase complex from Pyrococcus furiosus at 2.76 A

Summary for 5E0K

• Entry DOI: 10.2210/pdb5e0k/pdb
• Related: 5DVZ 5DW0 5DW3
• Descriptor: Tryptophan synthase alpha chain, Tryptophan synthase beta chain 1, PHOSPHATE ION, ... (4 entities in total)
• Functional Keywords: complex, plp, lyase, enzyme
• Biological source: Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1); More
• Total number of polymer chains: 12
• Total formula weight: 429095.17

Authors

Buller, A.R.,Murciano-Calles, J.,Arnold, F.H. (deposition date: 2015-09-29, release date: 2015-11-11, Last modification date: 2025-04-02)

Primary citation

Buller, A.R.,Brinkmann-Chen, S.,Romney, D.K.,Herger, M.,Murciano-Calles, J.,Arnold, F.H.
Directed evolution of the tryptophan synthase beta-subunit for stand-alone function recapitulates allosteric activation.
Proc.Natl.Acad.Sci.USA, 112:14599-14604, 2015

PubMed Abstract: 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 β-subunit of tryptophan synthase from Pyrococcus furiosus (PfTrpB). As part of its native αββα complex, TrpB efficiently produces tryptophan and tryptophan analogs; activity drops considerably when it is used as a stand-alone catalyst without the α-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 α-subunit. The engineered PfTrpB is a powerful platform for production of Trp analogs and for further directed evolution to expand substrate and reaction scope.

PubMed: 26553994
DOI: 10.1073/pnas.1516401112

Experimental method

X-RAY DIFFRACTION (2.76 Å)