Expression and purification of affinity-tagged variants of the photochemical reaction center from Heliobacterium modesticaldum

Orf, Gregory S.; Redding, Kevin E.

doi:10.1007/s11120-019-00672-3

Title: Expression and purification of affinity-tagged variants of the photochemical reaction center from Heliobacterium modesticaldum

Journal Article · 21 September 2019 · Photosynthesis Research

DOI: https://doi.org/10.1007/s11120-019-00672-3 · OSTI ID:1631822

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Arizona State Univ., Tempe, AZ (United States)

The heliobacterial photochemical reaction center (HbRC) from the chlorophototrophic Firmicutes bacterium Heliobacterium modesticaldum is the only homodimeric type I RC whose structure is known. Using genetic techniques recently established in our lab, we have developed a rapid heterologous expression system for the HbRC core polypeptide PshA. Our system relies on rescue of the non-chlorophototrophic ΔpshA::cbp2p-aph3 strain of Hbt. modesticaldum by expression of a heterologous pshA gene from a replicating shuttle vector. Furthermore, we constructed two tagged variants of PshA, one with an N-terminal octahistidine tag and one with an internal hexahistidine tag, which facilitate rapid purification of pure, active HbRC cores in milligram quantities. We constructed a suite of shuttle vectors bearing untagged or tagged versions of pshA driven by various promoters. Surprisingly, we found that the eno and gapDH_2 promoters from Clostridium thermocellum drive better expression of pshA than fragments of DNA derived from the region upstream of the pshA locus on the Hbt. modesticaldum genome. This “pshA rescue” strategy additionally provided a useful window into how Hbt. modesticaldum regulates pigment synthesis and growth rate when chlorophototrophic output decreases.

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Research Organization:: Arizona State Univ., Tempe, AZ (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences, and Biosciences Division

Grant/Contract Number:: SC0010575

OSTI ID:: 1631822

Journal Information:: Photosynthesis Research, Vol. 142, Issue 3; ISSN 0166-8595

Publisher:: SpringerCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 4 works

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References (38)

A heterogeneous tag-attachment to the homodimeric type 1 photosynthetic reaction center core protein in the green sulfur bacterium Chlorobaculum tepidum Azai, Chihiro; Kim, Kwang; Kondo, Toru Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1807, Issue 7 https://doi.org/10.1016/j.bbabio.2011.03.007	journal	July 2011
Prokaryotic photosynthesis and phototrophy illuminated Bryant, Donald A.; Frigaard, Niels-Ulrik Trends in Microbiology, Vol. 14, Issue 11 https://doi.org/10.1016/j.tim.2006.09.001	journal	November 2006
Silver staining of proteins in polyacrylamide gels Chevallet, Mireille; Luche, Sylvie; Rabilloud, Thierry Nature Protocols, Vol. 1, Issue 4 https://doi.org/10.1038/nprot.2006.288	journal	November 2006
Evolution of Oxygenic Photosynthesis Fischer, Woodward W.; Hemp, James; Johnson, Jena E. Annual Review of Earth and Planetary Sciences, Vol. 44, Issue 1 https://doi.org/10.1146/annurev-earth-060313-054810	journal	June 2016
Complete nucleotide sequence and genetic organization of the bacteriocinogenic plasmid, pIP404, from Clostridium perfringens Garnier, Thierry; Cole, Stewart T. Plasmid, Vol. 19, Issue 2 https://doi.org/10.1016/0147-619X(88)90052-2	journal	March 1988
Structure of a symmetric photosynthetic reaction center–photosystem Gisriel, Christopher; Sarrou, Iosifina; Ferlez, Bryan Science, Vol. 357, Issue 6355 https://doi.org/10.1126/science.aan5611	journal	July 2017
Rapid isolation of bacterial photosynthetic reaction centers with an engineered poly-histidine tag Goldsmith, Joshua O.; Boxer, Steven G. Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1276, Issue 3 https://doi.org/10.1016/0005-2728(96)00091-6	journal	September 1996
Purification of His6-tagged Photosystem I from Chlamydomonas reinhardtii Gulis, Galina; Narasimhulu, Kuppala V.; Fox, Lisa N. Photosynthesis Research, Vol. 96, Issue 1 https://doi.org/10.1007/s11120-007-9283-9	journal	January 2008
A modular system for Clostridium shuttle plasmids Heap, John T.; Pennington, Oliver J.; Cartman, Stephen T. Journal of Microbiological Methods, Vol. 78, Issue 1, p. 79-85 https://doi.org/10.1016/j.mimet.2009.05.004	journal	July 2009
Heliobacterial photosynthesis Heinnickel, Mark; Golbeck, John H. Photosynthesis Research, Vol. 92, Issue 1 https://doi.org/10.1007/s11120-007-9162-4	journal	April 2007
Resolution and Reconstitution of a Bound Fe−S Protein from the Photosynthetic Reaction Center of Heliobacterium modesticaldum ^† Heinnickel, Mark; Shen, Gaozhong; Agalarov, Rufat Biochemistry, Vol. 44, Issue 29 https://doi.org/10.1021/bi050588s	journal	July 2005
Identification and Characterization of PshB, the Dicluster Ferredoxin that Harbors the Terminal Electron Acceptors F _A and F _B in Heliobacterium modesticaldum ^† ^, ^‡ Heinnickel, Mark; Shen, Gaozhong; Golbeck, John H. Biochemistry, Vol. 46, Issue 9 https://doi.org/10.1021/bi0622165	journal	March 2007
Chlorophyll breakdown in higher plants Hörtensteiner, Stefan; Kräutler, Bernhard Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1807, Issue 8 https://doi.org/10.1016/j.bbabio.2010.12.007	journal	August 2011
Light-driven quinone reduction in heliobacterial membranes Kashey, Trevor S.; Luu, Dustin D.; Cowgill, John C. Photosynthesis Research, Vol. 138, Issue 1 https://doi.org/10.1007/s11120-018-0496-x	journal	March 2018
Heliobacterium modesticaldum, sp. nov., a thermophilic heliobacterium of hot springs and volcanic soils Kimble, Linda K.; Mandelco, Linda; Woese, Carl R. Archives of Microbiology, Vol. 163, Issue 4 https://doi.org/10.1007/BF00393378	journal	April 1995
Menaquinone as the Secondary Electron Acceptor in the Type I Homodimeric Photosynthetic Reaction Center of Heliobacterium modesticaldum Kondo, Toru; Itoh, Shigeru; Matsuoka, Masahiro The Journal of Physical Chemistry B, Vol. 119, Issue 27 https://doi.org/10.1021/acs.jpcb.5b03723	journal	June 2015
FastCloning: a highly simplified, purification-free, sequence- and ligation-independent PCR cloning method Li, Chaokun; Wen, Aiyun; Shen, Benchang BMC Biotechnology, Vol. 11, Issue 1 https://doi.org/10.1186/1472-6750-11-92	journal	January 2011
ESR Signal of the Iron−Sulfur Center F _X and Its Function in the Homodimeric Reaction Center of Heliobacterium modesticaldum ^† ^, ^‡ Miyamoto, Ryo; Iwaki, Masayo; Mino, Hiroyuki Biochemistry, Vol. 45, Issue 20 https://doi.org/10.1021/bi0519710	journal	May 2006
An Electron Spin-Polarized Signal of the P800 ⁺ A ₁ (Q) ⁻ State in the Homodimeric Reaction Center Core Complex of Heliobacterium modesticaldum ^† Miyamoto, Ryo; Mino, Hiroyuki; Kondo, Toru Biochemistry, Vol. 47, Issue 15 https://doi.org/10.1021/bi701612v	journal	April 2008
Identifying promoters for gene expression in Clostridium thermocellum Olson, Daniel G.; Maloney, Marybeth; Lanahan, Anthony A. Metabolic Engineering Communications, Vol. 2 https://doi.org/10.1016/j.meteno.2015.03.002	journal	December 2015
Evolution of photosynthetic reaction centers: insights from the structure of the heliobacterial reaction center Orf, Gregory S.; Gisriel, Christopher; Redding, Kevin E. Photosynthesis Research, Vol. 138, Issue 1 https://doi.org/10.1007/s11120-018-0503-2	journal	March 2018
Engineered biosynthesis of bacteriochlorophyll gF in Rhodobacter sphaeroides Ortega-Ramos, Marcia; Canniffe, Daniel P.; Radle, Matthew I. Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1859, Issue 7 https://doi.org/10.1016/j.bbabio.2018.02.006	journal	July 2018
Modulation of the fluorescence yield in heliobacterial cells by induction of charge recombination in the photosynthetic reaction center Redding, Kevin E.; Sarrou, Iosifina; Rappaport, Fabrice Photosynthesis Research, Vol. 120, Issue 1-2 https://doi.org/10.1007/s11120-013-9957-4	journal	December 2013
Purification of the photosynthetic reaction center from Heliobacterium modesticaldum Sarrou, Iosifina; Khan, Zahid; Cowgill, John Photosynthesis Research, Vol. 111, Issue 3 https://doi.org/10.1007/s11120-012-9726-9	journal	March 2012
Tricine–SDS-PAGE Schägger, Hermann Nature Protocols, Vol. 1, Issue 1 https://doi.org/10.1038/nprot.2006.4	journal	May 2006
Genome-wide Transcriptional Response during the Shift to N2-fixing Conditions in Heliobacterium modesticaldum Sheehy, Daniel; Kuang Lu, Yih; Osman, Fawsia Journal of Proteomics & Bioinformatics, Vol. 11, Issue 8 https://doi.org/10.4172/jpb.1000481	journal	January 2018
In-gel digestion for mass spectrometric characterization of proteins and proteomes Shevchenko, Andrej; Tomas, Henrik; Havli, Jan Nature Protocols, Vol. 1, Issue 6 https://doi.org/10.1038/nprot.2006.468	journal	December 2006
Arabidopsis STAY-GREEN , Mendel’s Green Cotyledon Gene, Encodes Magnesium-Dechelatase Shimoda, Yousuke; Ito, Hisashi; Tanaka, Ayumi The Plant Cell, Vol. 28, Issue 9 https://doi.org/10.1105/tpc.16.00428	journal	September 2016
High-performance liquid chromatography of chloroplast pigments Siefermann-Harms, Dorothea Journal of Chromatography A, Vol. 448 https://doi.org/10.1016/S0021-9673(01)84604-5	journal	January 1988
Construction of a sequencedClostridium perfringens-Escherichia coli shuttle plasmid Sloan, Joan; Warner, Tracy A.; Scott, Paul T. Plasmid, Vol. 27, Issue 3 https://doi.org/10.1016/0147-619X(92)90023-4	journal	May 1992
Highly Purified Thermo-Stable Oxygen-Evolving Photosystem II Core Complex from the Thermophilic Cyanobacterium Synechococcus elongatus Having His-Tagged CP43 Sugiura, Miwa; Inoue, Yorinao Plant and Cell Physiology, Vol. 40, Issue 12 https://doi.org/10.1093/oxfordjournals.pcp.a029510	journal	January 1999
Carbon Flow of Heliobacteria Is Related More to Clostridia than to the Green Sulfur Bacteria Tang, Kuo-Hsiang; Feng, Xueyang; Zhuang, Wei-Qin Journal of Biological Chemistry, Vol. 285, Issue 45 https://doi.org/10.1074/jbc.M110.163303	journal	August 2010
Isolation of a photoactive photosynthetic reaction center-core antenna complex from Heliobacillus mobilis Trost, Jeffrey T.; Blankenship, Robert E. Biochemistry, Vol. 28, Issue 26 https://doi.org/10.1021/bi00452a003	journal	December 1989
Purification and properties of an antenna-reaction center complex from heliobacteria van de Meent, Erik Jan; Kleinherenbrink, Frank A. M.; Amesz, Jan Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1015, Issue 2 https://doi.org/10.1016/0005-2728(90)90024-X	journal	February 1990
Identification of 81-hydroxychlorophyll a as a functional reaction center pigment in heliobacteria van de Meent, Erik Jan; Kobayashi, Masami; Erkelens, Cornelis Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1058, Issue 3 https://doi.org/10.1016/S0005-2728(05)80131-8	journal	July 1991
Transient electron paramagnetic resonance spectroscopy on green-sulfur bacteria and heliobacteria at two microwave frequencies van der Est, A.; Hager-Braun, C.; Leibl, W. Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1409, Issue 2 https://doi.org/10.1016/S0005-2728(98)00152-2	journal	December 1998
Heliobacterium modesticaldum, sp. nov., a thermophilic heliobacterium of hot springs and volcanic soils Kimble, Linda K.; Mandelco, Linda; Woese, Carl R. Archives of Microbiology, Vol. 163, Issue 4 https://doi.org/10.1007/s002030050202	journal	April 1995
Chlorophyll breakdown in higher plants. Hörtensteiner, S.; Kräutler, B. Elsevier https://doi.org/10.5167/uzh-53843	text	January 2011

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Title: Expression and purification of affinity-tagged variants of the photochemical reaction center from Heliobacterium modesticaldum

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