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Reaction centers of the thermophilic microaerophile, Chloracidobacterium thermophilum (Acidobacteria) I: biochemical and biophysical characterization

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Abstract

Chloracidobacterium thermophilum is a microaerophilic, anoxygenic member of the green chlorophototrophic bacteria. This bacterium is the first characterized oxygen-requiring chlorophototroph with chlorosomes, the FMO protein, and homodimeric type-1 reaction centers (RCs). The RCs of C. thermophilum are also unique because they contain three types of chlorophylls, bacteriochlorophyll aP esterified with phytol, Chl aPD esterified with Δ2,6-phytadienol, and Zn-BChl aP′ esterified with phytol, in the approximate molar ratio 32:24:4. The light-induced difference spectrum of these RCs had a bleaching maximum at 839 nm and also revealed an electrochromic bandshift that is probably derived from a BChl a molecule near P840+. The FX [4Fe–4S] cluster had a midpoint potential of ca. − 581 mV, and the spectroscopic properties of the P+ F −X spin-polarized radical pair were very similar to those of reaction centers of heliobacteria and green sulfur bacteria. The data further indicate that electron transfer occurs directly from A0− to FX, as occurs in other homodimeric type-1 RCs. Washing experiments with isolated membranes suggested that the PscB subunit of these reaction centers is more tightly bound than PshB in heliobacteria. Thus, the reaction centers of C. thermophilum have some properties that resemble other homodimeric reaction centers but also have specific properties that are more similar to those of Photosystem I. These differences probably contribute to protection of the electron transfer chain from oxygen, contributing to the oxygen tolerance of this microaerophile.

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Abbreviations

BChl:

Bacteriochlorophyll

BPheo:

Bacteriopheophytin

C :

Chloracidobacterium

Cba :

Chlorobaculum

Chl:

Chlorophyll

CW-EPR:

Continuous wave, field-modulated, time-resolved continuous-wave electron paramagnetic resonance

β-DDM:

n-dodecyl-β-maltoside

EPR:

Electron paramagnetic resonance

FMO:

Fenna–Matthews–Olson bacteriochlorophyll a-binding protein

HEPES:

4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid

HPLC:

High-performance liquid chromatography

2D-HYSCORE:

Two-dimensional hyperfine sublevel correlation spectroscopy

LC–MS-MS:

Liquid chromatography-tandem mass spectrometry

LED:

Light-emitting diode

OD:

Optical density

P:

Phyol

PAGE:

Polyacrylamide gel electrophoresis

PBS:

Phosphate-buffered saline

PD:

2,6-Phytadienol

Pheo:

Pheophytin

photo-CIDNP:

Photochemically induced dynamic nuclear polarization

PSI:

Photosystem I

PSII:

Photosystem II

RC(s):

Reaction center(s)

RP:

Reversed phase

SDS:

Sodium dodecyl sulfate

SHE:

Standard hydrogen electrode

trEPR:

Transient electron paramagnetic resonance

ZFS:

Zero field splitting

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Acknowledgements

The authors thank Dr. Tatiana N. Laremore of the Proteomics and Mass Spectrometry core facility (Huck Institutes for the Life Sciences, The Pennsylvania State University) for performing the tryptic peptide fingerprinting studies reported here. The authors would also like to thank Dr. Art van der Est for helpful discussions about transient EPR data. Studies in the laboratories of D.A.B. and J.H.G. were supported by Grants Nos. DE-FG02-94ER20137 and DE-SC0010575, respectively, from the Photosynthetic Systems Program, Division of Chemical Sciences, Geosciences, and Biosciences (CSGB), Office of Basic Energy Sciences of the U. S. Department of Energy.

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Author notes

  1. Bryan Ferlez

    Present address: MSU-DOE Plant Research Laboratory, Michigan State University, 612 Wilson Road, East Lansing, MI, 48824, USA

  2. Marcus Tank

    Present address: Department of Biological Sciences, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo, 192-0397, Japan

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, The Pennsylvania State University, S-002 Frear Building, University Park, PA, 16802, USA

    Zhihui He, Bryan Ferlez, Vasily Kurashov, Marcus Tank, John H. Golbeck & Donald A. Bryant

  2. Department of Chemistry, The Pennsylvania State University, University Park, PA, 16802, USA

    John H. Golbeck

  3. Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA

    Donald A. Bryant

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  1. Zhihui He
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Correspondence to Donald A. Bryant.

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He, Z., Ferlez, B., Kurashov, V. et al. Reaction centers of the thermophilic microaerophile, Chloracidobacterium thermophilum (Acidobacteria) I: biochemical and biophysical characterization. Photosynth Res 142, 87–103 (2019). https://doi.org/10.1007/s11120-019-00650-9

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