TY - JOUR
T1 - Variation in LOV Photoreceptor Activation Dynamics Probed by Time-Resolved Infrared Spectroscopy
AU - Iuliano, James N.
AU - Gil, Agnieszka A.
AU - Laptenok, Sergey P.
AU - Hall, Christopher R.
AU - Tolentino Collado, Jinnette
AU - Lukacs, Andras
AU - Hag Ahmed, Safaa A.
AU - Abyad, Jenna
AU - Daryaee, Taraneh
AU - Greetham, Gregory M.
AU - Sazanovich, Igor V.
AU - Illarionov, Boris
AU - Bacher, Adelbert
AU - Fischer, Markus
AU - Towrie, Michael
AU - French, Jarrod B.
AU - Meech, Stephen R.
AU - Tonge, Peter J.
N1 - Publisher Copyright:
© 2017 American Chemical Society.
PY - 2018/2/6
Y1 - 2018/2/6
N2 - The light, oxygen, voltage (LOV) domain proteins are blue light photoreceptors that utilize a noncovalently bound flavin mononucleotide (FMN) cofactor as the chromophore. The modular nature of these proteins has led to their wide adoption in the emerging fields of optogenetics and optobiology, where the LOV domain has been fused to a variety of output domains leading to novel light-controlled applications. In this work, we extend our studies of the subpicosecond to several hundred microsecond transient infrared spectroscopy of the isolated LOV domain AsLOV2 to three full-length photoreceptors in which the LOV domain is fused to an output domain: the LOV-STAS protein, YtvA, the LOV-HTH transcription factor, EL222, and the LOV-histidine kinase, LovK. Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate. However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain. While the rate of adduct formation varies by only 3.6-fold among the proteins, the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
AB - The light, oxygen, voltage (LOV) domain proteins are blue light photoreceptors that utilize a noncovalently bound flavin mononucleotide (FMN) cofactor as the chromophore. The modular nature of these proteins has led to their wide adoption in the emerging fields of optogenetics and optobiology, where the LOV domain has been fused to a variety of output domains leading to novel light-controlled applications. In this work, we extend our studies of the subpicosecond to several hundred microsecond transient infrared spectroscopy of the isolated LOV domain AsLOV2 to three full-length photoreceptors in which the LOV domain is fused to an output domain: the LOV-STAS protein, YtvA, the LOV-HTH transcription factor, EL222, and the LOV-histidine kinase, LovK. Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate. However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain. While the rate of adduct formation varies by only 3.6-fold among the proteins, the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
UR - https://www.scopus.com/pages/publications/85041462465
U2 - 10.1021/acs.biochem.7b01040
DO - 10.1021/acs.biochem.7b01040
M3 - Article
C2 - 29239168
AN - SCOPUS:85041462465
SN - 0006-2960
VL - 57
SP - 620
EP - 630
JO - Biochemistry
JF - Biochemistry
IS - 5
ER -