Scanning probe microscopy of bacterial red light photoreceptors

Fernando G. Tobias; Anna Gawedzka; Max S. Goldmeier; Alexandra C. Sakols; Emina A. Stojković; Stefan Tsonchev; Kenneth T. Nicholson

doi:10.1557/opl.2012.1006

Scanning probe microscopy of bacterial red light photoreceptors

Fernando G. Tobias, Anna Gawedzka, Max S. Goldmeier, Alexandra C. Sakols, Emina A. Stojković, Stefan Tsonchev, Kenneth T. Nicholson

Chemistry

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

1 Scopus citations

Abstract

Bacteriophytochromes (Bphs) are red-light photoreceptors found in photosynthetic and non-photosynthetic bacteria that have been engineered into infrared fluorescent protein markers. Bphs are composed of a photosensory module that is covalently linked to an effector/regulatory module, usually a histidine kinase (HK) domain. Light-induced, global structural changes are proposed to originate within the covalently attached biliverdin chromophore, a linear tetrapyrrole, and propagate through the protein. Bphs undergo reversible photoconversion between two distinct red and far-red light absorbing states, denoted Pr and Pfr respectively. For most Bphs, Pr is the dark-adapted state. The energy dissipated during Pr/Pfr photoconversion is proposed to directly impact the infrared fluorescence quantum yield. At this time, only structures of three different Bphs have been published, all of truncated proteins in their respective dark-adapted states. We have utilized scanning probe microscopy (SPM) to investigate the structure of intact Bphs in the light-adapted state in order to gain new insight into the mechanism of photoconversion and fluorescence. Scanning tunneling microscopy (STM) analysis of a pair of Bphs from photosynthetic bacterium R. palustris, RpBphP2 (P2) and RpBphP3 (P3) in their light-adapted states is presented in these proceedings. The concentration of the depositing protein has a key role in the molecular arrangements observed on the highly-ordered pyrolytic graphite (HOPG) surface. For example, at a high protein concentration, a hexagonal lattice of Bphs is observed by STM on a HOPG surface. Upon dilution, the photoreceptors self-organize into fiber-like structures on the surface. In these fibers, the dimer interface and the individual domains of the Bphs can be assigned and directly compared to a structural model of the intact, full-length proteins. In summary, SPM has potential to be an effective method for gaining new insight into Bph structure and dynamics.

Original language	English (US)
Title of host publication	Structure/Property Relationships in Biological and Biomimetic Materials at the Micro-, Nano- and Atomic-Length Scales
Pages	26-31
Number of pages	6
DOIs	https://doi.org/10.1557/opl.2012.1006
State	Published - 2012
Event	2012 MRS Spring Meeting - San Francisco, CA, United States Duration: Apr 9 2012 → Apr 13 2012

Publication series

Name	Materials Research Society Symposium Proceedings
Volume	1465
ISSN (Print)	0272-9172

Other

Other	2012 MRS Spring Meeting
Country/Territory	United States
City	San Francisco, CA
Period	4/9/12 → 4/13/12

Funding

We would like to thank the Northeastern Illinois University Student Center for Science Engagement for generous financial support (United States Department of Education CCRAA HIS Grant, Award # P031C080027). This support has provided student and faculty research stipends as well as monetary support for consumable supplies. We also acknowledge financial support from the NIH EARDA pilot grant: 5G11HD049644-04 to E.A.S.

ASJC Scopus subject areas

General Materials Science
Condensed Matter Physics
Mechanics of Materials
Mechanical Engineering

Access to Document

10.1557/opl.2012.1006

Cite this

Tobias, F. G., Gawedzka, A., Goldmeier, M. S., Sakols, A. C., Stojković, E. A., Tsonchev, S., & Nicholson, K. T. (2012). Scanning probe microscopy of bacterial red light photoreceptors. In Structure/Property Relationships in Biological and Biomimetic Materials at the Micro-, Nano- and Atomic-Length Scales (pp. 26-31). (Materials Research Society Symposium Proceedings; Vol. 1465). https://doi.org/10.1557/opl.2012.1006

@inproceedings{cfd8a215d1374ee482e94e590e858187,

title = "Scanning probe microscopy of bacterial red light photoreceptors",

abstract = "Bacteriophytochromes (Bphs) are red-light photoreceptors found in photosynthetic and non-photosynthetic bacteria that have been engineered into infrared fluorescent protein markers. Bphs are composed of a photosensory module that is covalently linked to an effector/regulatory module, usually a histidine kinase (HK) domain. Light-induced, global structural changes are proposed to originate within the covalently attached biliverdin chromophore, a linear tetrapyrrole, and propagate through the protein. Bphs undergo reversible photoconversion between two distinct red and far-red light absorbing states, denoted Pr and Pfr respectively. For most Bphs, Pr is the dark-adapted state. The energy dissipated during Pr/Pfr photoconversion is proposed to directly impact the infrared fluorescence quantum yield. At this time, only structures of three different Bphs have been published, all of truncated proteins in their respective dark-adapted states. We have utilized scanning probe microscopy (SPM) to investigate the structure of intact Bphs in the light-adapted state in order to gain new insight into the mechanism of photoconversion and fluorescence. Scanning tunneling microscopy (STM) analysis of a pair of Bphs from photosynthetic bacterium R. palustris, RpBphP2 (P2) and RpBphP3 (P3) in their light-adapted states is presented in these proceedings. The concentration of the depositing protein has a key role in the molecular arrangements observed on the highly-ordered pyrolytic graphite (HOPG) surface. For example, at a high protein concentration, a hexagonal lattice of Bphs is observed by STM on a HOPG surface. Upon dilution, the photoreceptors self-organize into fiber-like structures on the surface. In these fibers, the dimer interface and the individual domains of the Bphs can be assigned and directly compared to a structural model of the intact, full-length proteins. In summary, SPM has potential to be an effective method for gaining new insight into Bph structure and dynamics.",

author = "Tobias, {Fernando G.} and Anna Gawedzka and Goldmeier, {Max S.} and Sakols, {Alexandra C.} and Stojkovi{\'c}, {Emina A.} and Stefan Tsonchev and Nicholson, {Kenneth T.}",

note = "Funding Information: We would like to thank the Northeastern Illinois University Student Center for Science Engagement for generous financial support (United States Department of Education CCRAA HIS Grant, Award # P031C080027). This support has provided student and faculty research stipends as well as monetary support for consumable supplies. We also acknowledge financial support from the NIH EARDA pilot grant: 5G11HD049644-04 to E.A.S.; 2012 MRS Spring Meeting ; Conference date: 09-04-2012 Through 13-04-2012",

year = "2012",

doi = "10.1557/opl.2012.1006",

language = "English (US)",

isbn = "9781627482585",

series = "Materials Research Society Symposium Proceedings",

pages = "26--31",

booktitle = "Structure/Property Relationships in Biological and Biomimetic Materials at the Micro-, Nano- and Atomic-Length Scales",

}

Tobias, FG, Gawedzka, A, Goldmeier, MS, Sakols, AC, Stojković, EA, Tsonchev, S & Nicholson, KT 2012, Scanning probe microscopy of bacterial red light photoreceptors. in Structure/Property Relationships in Biological and Biomimetic Materials at the Micro-, Nano- and Atomic-Length Scales. Materials Research Society Symposium Proceedings, vol. 1465, pp. 26-31, 2012 MRS Spring Meeting, San Francisco, CA, United States, 4/9/12. https://doi.org/10.1557/opl.2012.1006

Scanning probe microscopy of bacterial red light photoreceptors. / Tobias, Fernando G.; Gawedzka, Anna; Goldmeier, Max S. et al.
Structure/Property Relationships in Biological and Biomimetic Materials at the Micro-, Nano- and Atomic-Length Scales. 2012. p. 26-31 (Materials Research Society Symposium Proceedings; Vol. 1465).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

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T1 - Scanning probe microscopy of bacterial red light photoreceptors

AU - Tobias, Fernando G.

AU - Gawedzka, Anna

AU - Goldmeier, Max S.

AU - Sakols, Alexandra C.

AU - Stojković, Emina A.

AU - Tsonchev, Stefan

AU - Nicholson, Kenneth T.

N1 - Funding Information: We would like to thank the Northeastern Illinois University Student Center for Science Engagement for generous financial support (United States Department of Education CCRAA HIS Grant, Award # P031C080027). This support has provided student and faculty research stipends as well as monetary support for consumable supplies. We also acknowledge financial support from the NIH EARDA pilot grant: 5G11HD049644-04 to E.A.S.

PY - 2012

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N2 - Bacteriophytochromes (Bphs) are red-light photoreceptors found in photosynthetic and non-photosynthetic bacteria that have been engineered into infrared fluorescent protein markers. Bphs are composed of a photosensory module that is covalently linked to an effector/regulatory module, usually a histidine kinase (HK) domain. Light-induced, global structural changes are proposed to originate within the covalently attached biliverdin chromophore, a linear tetrapyrrole, and propagate through the protein. Bphs undergo reversible photoconversion between two distinct red and far-red light absorbing states, denoted Pr and Pfr respectively. For most Bphs, Pr is the dark-adapted state. The energy dissipated during Pr/Pfr photoconversion is proposed to directly impact the infrared fluorescence quantum yield. At this time, only structures of three different Bphs have been published, all of truncated proteins in their respective dark-adapted states. We have utilized scanning probe microscopy (SPM) to investigate the structure of intact Bphs in the light-adapted state in order to gain new insight into the mechanism of photoconversion and fluorescence. Scanning tunneling microscopy (STM) analysis of a pair of Bphs from photosynthetic bacterium R. palustris, RpBphP2 (P2) and RpBphP3 (P3) in their light-adapted states is presented in these proceedings. The concentration of the depositing protein has a key role in the molecular arrangements observed on the highly-ordered pyrolytic graphite (HOPG) surface. For example, at a high protein concentration, a hexagonal lattice of Bphs is observed by STM on a HOPG surface. Upon dilution, the photoreceptors self-organize into fiber-like structures on the surface. In these fibers, the dimer interface and the individual domains of the Bphs can be assigned and directly compared to a structural model of the intact, full-length proteins. In summary, SPM has potential to be an effective method for gaining new insight into Bph structure and dynamics.

AB - Bacteriophytochromes (Bphs) are red-light photoreceptors found in photosynthetic and non-photosynthetic bacteria that have been engineered into infrared fluorescent protein markers. Bphs are composed of a photosensory module that is covalently linked to an effector/regulatory module, usually a histidine kinase (HK) domain. Light-induced, global structural changes are proposed to originate within the covalently attached biliverdin chromophore, a linear tetrapyrrole, and propagate through the protein. Bphs undergo reversible photoconversion between two distinct red and far-red light absorbing states, denoted Pr and Pfr respectively. For most Bphs, Pr is the dark-adapted state. The energy dissipated during Pr/Pfr photoconversion is proposed to directly impact the infrared fluorescence quantum yield. At this time, only structures of three different Bphs have been published, all of truncated proteins in their respective dark-adapted states. We have utilized scanning probe microscopy (SPM) to investigate the structure of intact Bphs in the light-adapted state in order to gain new insight into the mechanism of photoconversion and fluorescence. Scanning tunneling microscopy (STM) analysis of a pair of Bphs from photosynthetic bacterium R. palustris, RpBphP2 (P2) and RpBphP3 (P3) in their light-adapted states is presented in these proceedings. The concentration of the depositing protein has a key role in the molecular arrangements observed on the highly-ordered pyrolytic graphite (HOPG) surface. For example, at a high protein concentration, a hexagonal lattice of Bphs is observed by STM on a HOPG surface. Upon dilution, the photoreceptors self-organize into fiber-like structures on the surface. In these fibers, the dimer interface and the individual domains of the Bphs can be assigned and directly compared to a structural model of the intact, full-length proteins. In summary, SPM has potential to be an effective method for gaining new insight into Bph structure and dynamics.

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SN - 9781627482585

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EP - 31

BT - Structure/Property Relationships in Biological and Biomimetic Materials at the Micro-, Nano- and Atomic-Length Scales

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ER -

Scanning probe microscopy of bacterial red light photoreceptors

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