Modulating LOV domain photodynamics with a residue alteration outside the chromophore binding site

Sang Hun Song, Peter L. Freddolino, Abigail I. Nash, Elizabeth C. Carroll, Klaus Schulten, Kevin H. Gardner, Delmar S. Larsen

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Phototropins, a class of light-activated protein kinases, are essential for several blue light responses in plants and algae, including phototropism. These proteins contain two internal light, oxygen, and voltage sensitive (LOV) domains, which bind flavin chromophores and undergo a reversible photochemical formation of a cysteinyl-flavin adduct as part of the light sensing process. While the photodynamic properties of such photosensory domains are dictated by interactions between the chromophore and surrounding protein, more distant residues can play a significant role as well. Here we explore the role of the Phe434 residue in the photosensory response of the second LOV domain of Avena sativa phototropin 1 (AsLOV2), a model photochemical system for these LOV domains. Phe434 is more than 6 Å from the FMN chromophore in AsLOV2; nevertheless, an F434Y point mutation is likely to change several structural features of the chromophore binding site, as we demonstrate using molecular dynamics simulations. Transient absorption signals spanning 15 decades in time were compared for wild-type AsLOV2 and the F434Y mutant, showing that the latter has significantly altered photodynamics, including (i) a faster intersystem crossing leading to triplet formation on a nanosecond time scale, (ii) biphasic formation of adduct-state kinetics on the microsecond time scale, and (iii) greatly accelerated ground-state recovery kinetics on a second time scale. We present mechanistic models that link these spectroscopic differences to changes in the configuration of the critical cysteine residue and in the chromophore's accessibility to solvent and oxygen according to MD trajectories and purging experiments. Taken together, these results demonstrate the importance of residues outside the chromophore-binding pocket in modulating LOV domain photodynamics.

Original languageEnglish (US)
Pages (from-to)2411-2423
Number of pages13
JournalBiochemistry
Volume50
Issue number13
DOIs
StatePublished - Apr 5 2011

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Chromophores
Binding Sites
Oxygen
Light
Electric potential
Phototropins
Phototropism
Flavin Mononucleotide
Purging
Kinetics
Molecular Dynamics Simulation
Algae
Point Mutation
Protein Kinases
Ground state
Cysteine
Molecular dynamics
Proteins
Trajectories
Recovery

ASJC Scopus subject areas

  • Biochemistry

Cite this

Song, S. H., Freddolino, P. L., Nash, A. I., Carroll, E. C., Schulten, K., Gardner, K. H., & Larsen, D. S. (2011). Modulating LOV domain photodynamics with a residue alteration outside the chromophore binding site. Biochemistry, 50(13), 2411-2423. https://doi.org/10.1021/bi200198x

Modulating LOV domain photodynamics with a residue alteration outside the chromophore binding site. / Song, Sang Hun; Freddolino, Peter L.; Nash, Abigail I.; Carroll, Elizabeth C.; Schulten, Klaus; Gardner, Kevin H.; Larsen, Delmar S.

In: Biochemistry, Vol. 50, No. 13, 05.04.2011, p. 2411-2423.

Research output: Contribution to journalArticle

Song, SH, Freddolino, PL, Nash, AI, Carroll, EC, Schulten, K, Gardner, KH & Larsen, DS 2011, 'Modulating LOV domain photodynamics with a residue alteration outside the chromophore binding site', Biochemistry, vol. 50, no. 13, pp. 2411-2423. https://doi.org/10.1021/bi200198x
Song, Sang Hun ; Freddolino, Peter L. ; Nash, Abigail I. ; Carroll, Elizabeth C. ; Schulten, Klaus ; Gardner, Kevin H. ; Larsen, Delmar S. / Modulating LOV domain photodynamics with a residue alteration outside the chromophore binding site. In: Biochemistry. 2011 ; Vol. 50, No. 13. pp. 2411-2423.
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