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Directed evolution of the tryptophan synthase β-subunit for stand-alone function recapitulates allosteric activation.

Buller AR, Brinkmann-Chen S, Romney DK, Herger M, Murciano-Calles J, Arnold FH
Proc Natl Acad Sci U S A 112 14599-604 (2015)
Related entries: 5dvz, 5dw0, 5e0k

Cited: 69 times
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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.

Articles - 5dw3 mentioned but not cited (3)

  1. Directed evolution of the tryptophan synthase β-subunit for stand-alone function recapitulates allosteric activation. Buller AR, Brinkmann-Chen S, Romney DK, Herger M, Murciano-Calles J, Arnold FH. Proc Natl Acad Sci U S A 112 14599-14604 (2015)
  2. Tryptophan Synthase Uses an Atypical Mechanism To Achieve Substrate Specificity. Buller AR, van Roye P, Murciano-Calles J, Arnold FH. Biochemistry 55 7043-7046 (2016)
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Reviews citing this publication (13)

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Articles citing this publication (53)

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