TY - JOUR
T1 - Confinement in crystal lattice alters entire photocycle pathway of the Photoactive Yellow Protein
AU - Konold, Patrick E.
AU - Arik, Enis
AU - Weißenborn, Jörn
AU - Arents, Jos C.
AU - Hellingwerf, Klaas J.
AU - van Stokkum, Ivo H.M.
AU - Kennis, John T.M.
AU - Groot, Marie Louise
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Femtosecond time-resolved crystallography (TRC) on proteins enables resolving the spatial structure of short-lived photocycle intermediates. An open question is whether confinement and lower hydration of the proteins in the crystalline state affect the light-induced structural transformations. Here, we measured the full photocycle dynamics of a signal transduction protein often used as model system in TRC, Photoactive Yellow Protein (PYP), in the crystalline state and compared those to the dynamics in solution, utilizing electronic and vibrational transient absorption measurements from 100 fs over 12 decades in time. We find that the photocycle kinetics and structural dynamics of PYP in the crystalline form deviate from those in solution from the very first steps following photon absorption. This illustrates that ultrafast TRC results cannot be uncritically extrapolated to in vivo function, and that comparative spectroscopic experiments on proteins in crystalline and solution states can help identify structural intermediates under native conditions.
AB - Femtosecond time-resolved crystallography (TRC) on proteins enables resolving the spatial structure of short-lived photocycle intermediates. An open question is whether confinement and lower hydration of the proteins in the crystalline state affect the light-induced structural transformations. Here, we measured the full photocycle dynamics of a signal transduction protein often used as model system in TRC, Photoactive Yellow Protein (PYP), in the crystalline state and compared those to the dynamics in solution, utilizing electronic and vibrational transient absorption measurements from 100 fs over 12 decades in time. We find that the photocycle kinetics and structural dynamics of PYP in the crystalline form deviate from those in solution from the very first steps following photon absorption. This illustrates that ultrafast TRC results cannot be uncritically extrapolated to in vivo function, and that comparative spectroscopic experiments on proteins in crystalline and solution states can help identify structural intermediates under native conditions.
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U2 - 10.1038/s41467-020-18065-9
DO - 10.1038/s41467-020-18065-9
M3 - Article
C2 - 32843623
AN - SCOPUS:85089820479
SN - 2041-1723
VL - 11
SP - 1
EP - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 4248
ER -