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Daniel Otzen

In situ Sub-Cellular Identification of Functional Amyloids in Bacteria and Archaea by Infrared Nanospectroscopy

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In situ Sub-Cellular Identification of Functional Amyloids in Bacteria and Archaea by Infrared Nanospectroscopy. / Otzen, Daniel E.; Dueholm, Morten S.; Najarzadeh, Zahra; Knowles, Tuomas P.J.; Ruggeri, Francesco Simone.

In: Small Methods, Vol. 5, No. 6, 2001002, 06.2021.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Otzen, DE, Dueholm, MS, Najarzadeh, Z, Knowles, TPJ & Ruggeri, FS 2021, 'In situ Sub-Cellular Identification of Functional Amyloids in Bacteria and Archaea by Infrared Nanospectroscopy', Small Methods, vol. 5, no. 6, 2001002. https://doi.org/10.1002/smtd.202001002

APA

Otzen, D. E., Dueholm, M. S., Najarzadeh, Z., Knowles, T. P. J., & Ruggeri, F. S. (2021). In situ Sub-Cellular Identification of Functional Amyloids in Bacteria and Archaea by Infrared Nanospectroscopy. Small Methods, 5(6), [2001002]. https://doi.org/10.1002/smtd.202001002

CBE

MLA

Vancouver

Author

Otzen, Daniel E. ; Dueholm, Morten S. ; Najarzadeh, Zahra ; Knowles, Tuomas P.J. ; Ruggeri, Francesco Simone. / In situ Sub-Cellular Identification of Functional Amyloids in Bacteria and Archaea by Infrared Nanospectroscopy. In: Small Methods. 2021 ; Vol. 5, No. 6.

Bibtex

@article{3de3e7ef5fd94ea3b1d7e444135f4c62,
title = "In situ Sub-Cellular Identification of Functional Amyloids in Bacteria and Archaea by Infrared Nanospectroscopy",
abstract = "Formation of amyloid structures is originally linked to human disease. However, amyloid materials are found extensively in the animal and bacterial world where they stabilize intra- and extra-cellular environments like biofilms or cell envelopes. To date, functional amyloids have largely been studied using optical microscopy techniques in vivo, or after removal from their biological context for higher-resolution studies in vitro. Furthermore, conventional microscopies only indirectly identify amyloids based on morphology or unspecific amyloid dyes. Here, the high chemical and spatial (≈20 nm) resolution of Infrared Nanospectroscopy (AFM-IR) to investigate functional amyloid from Escherichia coli (curli), Pseudomonas (Fap), and the Archaea Methanosaeta (MspA) in situ is exploited. It is demonstrated that AFM-IR identifies amyloid protein within single intact cells through their cross β-sheet secondary structure, which has a unique spectroscopic signature in the amide I band of protein. Using this approach, nanoscale-resolved chemical images and spectra of purified curli and Methanosaeta cell wall sheaths are provided. The results highlight significant differences in secondary structure between E. coli cells with and without curli. Taken together, these results suggest that AFM-IR is a new and powerful label-free tool for in situ investigations of the biophysical state of functional amyloid and biomolecules in general.",
keywords = "AFM-IR, archaeal cell wall sheaths, curli, functional amyloids",
author = "Otzen, {Daniel E.} and Dueholm, {Morten S.} and Zahra Najarzadeh and Knowles, {Tuomas P.J.} and Ruggeri, {Francesco Simone}",
note = "Publisher Copyright: {\textcopyright} 2021 The Authors. Small Methods published by Wiley-VCH GmbH",
year = "2021",
month = jun,
doi = "10.1002/smtd.202001002",
language = "English",
volume = "5",
journal = "Small Methods",
issn = "2366-9608",
publisher = "WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim",
number = "6",

}

RIS

TY - JOUR

T1 - In situ Sub-Cellular Identification of Functional Amyloids in Bacteria and Archaea by Infrared Nanospectroscopy

AU - Otzen, Daniel E.

AU - Dueholm, Morten S.

AU - Najarzadeh, Zahra

AU - Knowles, Tuomas P.J.

AU - Ruggeri, Francesco Simone

N1 - Publisher Copyright: © 2021 The Authors. Small Methods published by Wiley-VCH GmbH

PY - 2021/6

Y1 - 2021/6

N2 - Formation of amyloid structures is originally linked to human disease. However, amyloid materials are found extensively in the animal and bacterial world where they stabilize intra- and extra-cellular environments like biofilms or cell envelopes. To date, functional amyloids have largely been studied using optical microscopy techniques in vivo, or after removal from their biological context for higher-resolution studies in vitro. Furthermore, conventional microscopies only indirectly identify amyloids based on morphology or unspecific amyloid dyes. Here, the high chemical and spatial (≈20 nm) resolution of Infrared Nanospectroscopy (AFM-IR) to investigate functional amyloid from Escherichia coli (curli), Pseudomonas (Fap), and the Archaea Methanosaeta (MspA) in situ is exploited. It is demonstrated that AFM-IR identifies amyloid protein within single intact cells through their cross β-sheet secondary structure, which has a unique spectroscopic signature in the amide I band of protein. Using this approach, nanoscale-resolved chemical images and spectra of purified curli and Methanosaeta cell wall sheaths are provided. The results highlight significant differences in secondary structure between E. coli cells with and without curli. Taken together, these results suggest that AFM-IR is a new and powerful label-free tool for in situ investigations of the biophysical state of functional amyloid and biomolecules in general.

AB - Formation of amyloid structures is originally linked to human disease. However, amyloid materials are found extensively in the animal and bacterial world where they stabilize intra- and extra-cellular environments like biofilms or cell envelopes. To date, functional amyloids have largely been studied using optical microscopy techniques in vivo, or after removal from their biological context for higher-resolution studies in vitro. Furthermore, conventional microscopies only indirectly identify amyloids based on morphology or unspecific amyloid dyes. Here, the high chemical and spatial (≈20 nm) resolution of Infrared Nanospectroscopy (AFM-IR) to investigate functional amyloid from Escherichia coli (curli), Pseudomonas (Fap), and the Archaea Methanosaeta (MspA) in situ is exploited. It is demonstrated that AFM-IR identifies amyloid protein within single intact cells through their cross β-sheet secondary structure, which has a unique spectroscopic signature in the amide I band of protein. Using this approach, nanoscale-resolved chemical images and spectra of purified curli and Methanosaeta cell wall sheaths are provided. The results highlight significant differences in secondary structure between E. coli cells with and without curli. Taken together, these results suggest that AFM-IR is a new and powerful label-free tool for in situ investigations of the biophysical state of functional amyloid and biomolecules in general.

KW - AFM-IR

KW - archaeal cell wall sheaths

KW - curli

KW - functional amyloids

UR - http://www.scopus.com/inward/record.url?scp=85105200950&partnerID=8YFLogxK

U2 - 10.1002/smtd.202001002

DO - 10.1002/smtd.202001002

M3 - Journal article

C2 - 34927901

AN - SCOPUS:85105200950

VL - 5

JO - Small Methods

JF - Small Methods

SN - 2366-9608

IS - 6

M1 - 2001002

ER -