Single-molecule FRET imaging of GPCR dimers in living cells

Research output: Contribution to journal › Journal article › Research › peer-review

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Single-molecule FRET imaging of GPCR dimers in living cells. / Asher, Wesley B.; Geggier, Peter; Holsey, Michael D.; Gilmore, Grant T.; Pati, Avik K.; Meszaros, Jozsef; Terry, Daniel S.; Mathiasen, Signe; Kaliszewski, Megan J.; McCauley, Mitchell D.; Govindaraju, Alekhya; Zhou, Zhou; Harikumar, Kaleeckal G.; Jaqaman, Khuloud; Miller, Laurence J; Smith, Adam W.; Blanchard, Scott C.; Javitch, Jonathan A.

In: Nature Methods, Vol. 18, No. 4, 2021, p. 397-405.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Asher, WB, Geggier, P, Holsey, MD, Gilmore, GT, Pati, AK, Meszaros, J, Terry, DS, Mathiasen, S, Kaliszewski, MJ, McCauley, MD, Govindaraju, A, Zhou, Z, Harikumar, KG, Jaqaman, K, Miller, LJ, Smith, AW, Blanchard, SC & Javitch, JA 2021, 'Single-molecule FRET imaging of GPCR dimers in living cells', Nature Methods, vol. 18, no. 4, pp. 397-405. https://doi.org/10.1038/s41592-021-01081-y

APA

Asher, W. B., Geggier, P., Holsey, M. D., Gilmore, G. T., Pati, A. K., Meszaros, J., Terry, D. S., Mathiasen, S., Kaliszewski, M. J., McCauley, M. D., Govindaraju, A., Zhou, Z., Harikumar, K. G., Jaqaman, K., Miller, L. J., Smith, A. W., Blanchard, S. C., & Javitch, J. A. (2021). Single-molecule FRET imaging of GPCR dimers in living cells. Nature Methods, 18(4), 397-405. https://doi.org/10.1038/s41592-021-01081-y

Vancouver

Asher WB, Geggier P, Holsey MD, Gilmore GT, Pati AK, Meszaros J et al. Single-molecule FRET imaging of GPCR dimers in living cells. Nature Methods. 2021;18(4):397-405. https://doi.org/10.1038/s41592-021-01081-y

Author

Asher, Wesley B. ; Geggier, Peter ; Holsey, Michael D. ; Gilmore, Grant T. ; Pati, Avik K. ; Meszaros, Jozsef ; Terry, Daniel S. ; Mathiasen, Signe ; Kaliszewski, Megan J. ; McCauley, Mitchell D. ; Govindaraju, Alekhya ; Zhou, Zhou ; Harikumar, Kaleeckal G. ; Jaqaman, Khuloud ; Miller, Laurence J ; Smith, Adam W. ; Blanchard, Scott C. ; Javitch, Jonathan A. / Single-molecule FRET imaging of GPCR dimers in living cells. In: Nature Methods. 2021 ; Vol. 18, No. 4. pp. 397-405.

Bibtex

@article{2b43550a314245b09a03db4488ca6737,
title = "Single-molecule FRET imaging of GPCR dimers in living cells",
abstract = "Class C G protein-coupled receptors (GPCRs) are known to form stable homodimers or heterodimers critical for function, but the oligomeric status of class A and B receptors, which constitute >90% of all GPCRs, remains hotly debated. Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful approach with the potential to reveal valuable insights into GPCR organization but has rarely been used in living cells to study protein systems. Here, we report generally applicable methods for using smFRET to detect and track transmembrane proteins diffusing within the plasma membrane of mammalian cells. We leverage this in-cell smFRET approach to show agonist-induced structural dynamics within individual metabotropic glutamate receptor dimers. We apply these methods to representative class A, B and C receptors, finding evidence for receptor monomers, density-dependent dimers and constitutive dimers, respectively.",
author = "Asher, {Wesley B.} and Peter Geggier and Holsey, {Michael D.} and Gilmore, {Grant T.} and Pati, {Avik K.} and Jozsef Meszaros and Terry, {Daniel S.} and Signe Mathiasen and Kaliszewski, {Megan J.} and McCauley, {Mitchell D.} and Alekhya Govindaraju and Zhou Zhou and Harikumar, {Kaleeckal G.} and Khuloud Jaqaman and Miller, {Laurence J} and Smith, {Adam W.} and Blanchard, {Scott C.} and Javitch, {Jonathan A.}",
note = "Publisher Copyright: {\textcopyright} 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2021",
doi = "10.1038/s41592-021-01081-y",
language = "English",
volume = "18",
pages = "397--405",
journal = "Nature Methods",
issn = "1548-7091",
publisher = "nature publishing group",
number = "4",

}

RIS

TY - JOUR

T1 - Single-molecule FRET imaging of GPCR dimers in living cells

AU - Asher, Wesley B.

AU - Geggier, Peter

AU - Holsey, Michael D.

AU - Gilmore, Grant T.

AU - Pati, Avik K.

AU - Meszaros, Jozsef

AU - Terry, Daniel S.

AU - Mathiasen, Signe

AU - Kaliszewski, Megan J.

AU - McCauley, Mitchell D.

AU - Govindaraju, Alekhya

AU - Zhou, Zhou

AU - Harikumar, Kaleeckal G.

AU - Jaqaman, Khuloud

AU - Miller, Laurence J

AU - Smith, Adam W.

AU - Blanchard, Scott C.

AU - Javitch, Jonathan A.

N1 - Publisher Copyright: © 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.

PY - 2021

Y1 - 2021

N2 - Class C G protein-coupled receptors (GPCRs) are known to form stable homodimers or heterodimers critical for function, but the oligomeric status of class A and B receptors, which constitute >90% of all GPCRs, remains hotly debated. Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful approach with the potential to reveal valuable insights into GPCR organization but has rarely been used in living cells to study protein systems. Here, we report generally applicable methods for using smFRET to detect and track transmembrane proteins diffusing within the plasma membrane of mammalian cells. We leverage this in-cell smFRET approach to show agonist-induced structural dynamics within individual metabotropic glutamate receptor dimers. We apply these methods to representative class A, B and C receptors, finding evidence for receptor monomers, density-dependent dimers and constitutive dimers, respectively.

AB - Class C G protein-coupled receptors (GPCRs) are known to form stable homodimers or heterodimers critical for function, but the oligomeric status of class A and B receptors, which constitute >90% of all GPCRs, remains hotly debated. Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful approach with the potential to reveal valuable insights into GPCR organization but has rarely been used in living cells to study protein systems. Here, we report generally applicable methods for using smFRET to detect and track transmembrane proteins diffusing within the plasma membrane of mammalian cells. We leverage this in-cell smFRET approach to show agonist-induced structural dynamics within individual metabotropic glutamate receptor dimers. We apply these methods to representative class A, B and C receptors, finding evidence for receptor monomers, density-dependent dimers and constitutive dimers, respectively.

U2 - 10.1038/s41592-021-01081-y

DO - 10.1038/s41592-021-01081-y

M3 - Journal article

C2 - 33686301

AN - SCOPUS:85102284460

VL - 18

SP - 397

EP - 405

JO - Nature Methods

JF - Nature Methods

SN - 1548-7091

IS - 4

ER -

ID: 311723370