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Low-frequency resonance Raman studies of the H(M202)G cavity mutant of bacterial photosynthetic reaction centers

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Abstract

Low-frequency (90–435 cm−1) NIR-excitation (875–900 nm) resonance Raman (RR) studies are reported for the H(M202)G cavity mutant of bacterial photosynthetic reaction centers (RCs) from Rb. sphaeroides that was first described by Goldsmith et al. [(1996) Biochemistry 35: 2421–2428]. In this mutant, the His residue that axially ligates the Mg ion of the M-side bacteriochlorophyll (BChl) of the special pair primary donor (P) is replaced by a non-ligating Gly residue. Regardless, the Mg ion of PM in the H(M202)G RCs remains pentacoordinates and is presumably ligated by a water molecule, although this axial ligand has not been definitively identified. The low-frequency RR studies of the H(M202)G RCs are accompanied by studies of RCs exchanged with D2O and incubated with imidazole (Im). The RR studies of the cavity mutant RCs reveal the following: (1) The structure of PM in the H(M202)G RCs is different from that of the wild-type, consistent with an altered BChl core. (2) A water ligand for PM in the H(M202)G RCs is generally consistent with the low-frequency RR spectra. The Mg-OH2 stretching vibration is tentatively assigned to a band at 318 cm−1, a frequency higher than that of the Mg-His stretch of the native pigment (∼ ∼235 cm−1). (3) The BChl core structure of PM in the cavity mutant is rendered similar (but not identical) to that of the wild-type when the adventitious water axial ligand is replaced by Im. (4) Exchange with D2O results in more global structural changes, likely involving the protein, which in turn affect the structure of the BChls in P. (5) Assignment of the low-frequency vibrational spectrum of P is generally more complex than originally suggested.

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Abbreviations

BChl:

bacteriochlorophyll

BPh:

bacteriopheophytin

DFT:

density functional theory

LDAO:

N,N-dimethyldodecylamine-N-oxide

P:

special pair dimer of BChls

Q:

quinone

RC:

reaction center

RR:

resonance Raman

tris:

tris[hydroxymethyl]aminomethane

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Acknowledgements

We thank Professor S. G. Boxer for providing the cultures of the cavity mutants and P. Dolan and A. deWinter for preparation of the RCs. This work was supported by Grants GM-39781 (D.F.B.) and GM-30353 (H.A.F.) from the National Institute of General Medical Sciences.

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Authors and Affiliations

  1. Department of Chemistry, University of California, Riverside, California, 92521-0403, USA

    Kazimierz Czarnecki, Lei Chen, James R. Diers &  David F. Bocian

  2. Department of Chemistry, University of Connecticut, 55 North Eagleville Road, Storrs, CT, 06269-3060, USA

    Harry A. Frank

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  1. Kazimierz Czarnecki
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  2. Lei Chen
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  3. James R. Diers
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  4. Harry A. Frank
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  5. David F. Bocian
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Correspondence to David F. Bocian.

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Czarnecki, K., Chen, L., Diers, J.R. et al. Low-frequency resonance Raman studies of the H(M202)G cavity mutant of bacterial photosynthetic reaction centers. Photosynth Res 88, 31–41 (2006). https://doi.org/10.1007/s11120-005-9019-7

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