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Structural and dynamic changes of photoactive yellow protein during its photocycle in solution

Nature Structural Biology volume 5, pages 568–570 (1998)Cite this article

Abstract

Light irradiation of photoactive yellow protein (PYP) induces a photocycle, in which red-shifted (pR) and blue-shifted (pB) intermediates have been characterized. An NMR study of the long-lived pB intermediate now reveals that it exhibits a large degree of disorder and exists as a family of multiple conformers that exchange on a millisecond time scale. This shows that the behavior of PYP in solution is different from what has been observed in the crystalline state. Furthermore, differential refolding to ground state pG is observed, whereby the central β-sheet and parts of the helical structure are formed first and the region around the chromophore at a later stage.

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Figure 1: a, Photocycle of PYP in solution as described in ref. 10.
Figure 2: Schematic outline of the NMR pulse sequences a, used for recording interleaved spectra of ground state (pG) and blue-shifted intermediate (pB) of PYP and b, for the measurement of build-up rates of pG cross-peaks.
Figure 3
Figure 4: Ribbon diagram of the energy-minimized average solution structure of the pG state13 (created with MOLMOL18).

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Acknowledgements

We thank A. Kroon and R. Cordfunke for providing the 15N enriched NMR sample and N. Zwanenburg for technical support during the laser setup. G.R. acknowledges a fellowship from Rhone-Poulenc.

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Author notes

  1. Geerten W. Vuister

    Present address: Department of Biophysical Chemistry, University of Nijmegen, Toemooiveld 1, 6525 ED, Nijmegen, The Netherlands

Authors and Affiliations

  1. Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, Utrecht, 3584 CH, The Netherlands

    Gilles Rubinstenn, Geerten W. Vuister, Frans A. A. Mulder, Petra E. Düx, Rolf Boelens & Robert Kaptein

  2. Laboratory for Microbiology, E. C. Slater Institute, University of Amsterdam, Nieuwe Achtergracht 127, Amsterdam, 1018 WS, The Netherlands

    Klaas J. Hellingwerf

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Correspondence to Robert Kaptein.

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Rubinstenn, G., Vuister, G., Mulder, F. et al. Structural and dynamic changes of photoactive yellow protein during its photocycle in solution. Nat Struct Mol Biol 5, 568–570 (1998). https://doi.org/10.1038/823

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