Affinity proteomics for interactome and phosphoproteome screening in synaptosomes

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Standard

Affinity proteomics for interactome and phosphoproteome screening in synaptosomes. / Engholm-Keller, Kasper; Bache, Nicolai; Rao, Sushma R.; Wark, Jesse R.; Larsen, Martin R.; Robinson, Phillip J.; Graham, Mark E.

Synaptosomes. ed. / Kathryn M. Murphy. Springer, 2018. p. 165–191 (Neuromethods, Vol. 141).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Harvard

Engholm-Keller, K, Bache, N, Rao, SR, Wark, JR, Larsen, MR, Robinson, PJ & Graham, ME 2018, Affinity proteomics for interactome and phosphoproteome screening in synaptosomes. in KM Murphy (ed.), Synaptosomes. Springer, Neuromethods, vol. 141, pp. 165–191. https://doi.org/10.1007/978-1-4939-8739-9_10

APA

Engholm-Keller, K., Bache, N., Rao, S. R., Wark, J. R., Larsen, M. R., Robinson, P. J., & Graham, M. E. (2018). Affinity proteomics for interactome and phosphoproteome screening in synaptosomes. In K. M. Murphy (Ed.), Synaptosomes (pp. 165–191). Springer. Neuromethods Vol. 141 https://doi.org/10.1007/978-1-4939-8739-9_10

Vancouver

Engholm-Keller K, Bache N, Rao SR, Wark JR, Larsen MR, Robinson PJ et al. Affinity proteomics for interactome and phosphoproteome screening in synaptosomes. In Murphy KM, editor, Synaptosomes. Springer. 2018. p. 165–191. (Neuromethods, Vol. 141). https://doi.org/10.1007/978-1-4939-8739-9_10

Author

Engholm-Keller, Kasper ; Bache, Nicolai ; Rao, Sushma R. ; Wark, Jesse R. ; Larsen, Martin R. ; Robinson, Phillip J. ; Graham, Mark E. / Affinity proteomics for interactome and phosphoproteome screening in synaptosomes. Synaptosomes. editor / Kathryn M. Murphy. Springer, 2018. pp. 165–191 (Neuromethods, Vol. 141).

Bibtex

@inbook{e8ddd6fd371846958bb89789137efa9d,
title = "Affinity proteomics for interactome and phosphoproteome screening in synaptosomes",
abstract = "Phosphorylation is an essential regulatory protein modification, which modulates many presynaptic processes, including synaptic vesicle trafficking. Protein phosphorylation impacts neurotransmitter release and is likely to be mechanistically important for presynaptic plasticity, although only a few mechanisms have been described to date. Thus, interactomics and phosphoproteomics are important tools to determine presynaptic mechanisms. Over the last two decades, mass spectrometry-based proteomics has become the method of choice for protein-protein and protein phosphorylation analysis. This protocol is based on wellestablished procedures in our laboratory and will describe the application of affinity purification of presynaptic proteins via GST-fusion protein pull-downs for the study of presynaptic phospho-signalling. The pull-down is followed by on-bead proteolysis, chemical stable isotope labelling for precise quantification, enrichment of phosphorylated peptides using TiO2, and LC-MS/MS. The method outlines the software used for peptide/protein identification and quantification, as well as some of the bioinformatics tools that are available for putting the regulated phosphorylation sites into a functional context.",
author = "Kasper Engholm-Keller and Nicolai Bache and Rao, {Sushma R.} and Wark, {Jesse R.} and Larsen, {Martin R.} and Robinson, {Phillip J.} and Graham, {Mark E.}",
year = "2018",
doi = "10.1007/978-1-4939-8739-9_10",
language = "English",
isbn = "978-1-4939-9379-6",
series = "Neuromethods",
publisher = "Springer",
pages = "165–191",
editor = "Murphy, {Kathryn M.}",
booktitle = "Synaptosomes",
address = "Switzerland",

}

RIS

TY - CHAP

T1 - Affinity proteomics for interactome and phosphoproteome screening in synaptosomes

AU - Engholm-Keller, Kasper

AU - Bache, Nicolai

AU - Rao, Sushma R.

AU - Wark, Jesse R.

AU - Larsen, Martin R.

AU - Robinson, Phillip J.

AU - Graham, Mark E.

PY - 2018

Y1 - 2018

N2 - Phosphorylation is an essential regulatory protein modification, which modulates many presynaptic processes, including synaptic vesicle trafficking. Protein phosphorylation impacts neurotransmitter release and is likely to be mechanistically important for presynaptic plasticity, although only a few mechanisms have been described to date. Thus, interactomics and phosphoproteomics are important tools to determine presynaptic mechanisms. Over the last two decades, mass spectrometry-based proteomics has become the method of choice for protein-protein and protein phosphorylation analysis. This protocol is based on wellestablished procedures in our laboratory and will describe the application of affinity purification of presynaptic proteins via GST-fusion protein pull-downs for the study of presynaptic phospho-signalling. The pull-down is followed by on-bead proteolysis, chemical stable isotope labelling for precise quantification, enrichment of phosphorylated peptides using TiO2, and LC-MS/MS. The method outlines the software used for peptide/protein identification and quantification, as well as some of the bioinformatics tools that are available for putting the regulated phosphorylation sites into a functional context.

AB - Phosphorylation is an essential regulatory protein modification, which modulates many presynaptic processes, including synaptic vesicle trafficking. Protein phosphorylation impacts neurotransmitter release and is likely to be mechanistically important for presynaptic plasticity, although only a few mechanisms have been described to date. Thus, interactomics and phosphoproteomics are important tools to determine presynaptic mechanisms. Over the last two decades, mass spectrometry-based proteomics has become the method of choice for protein-protein and protein phosphorylation analysis. This protocol is based on wellestablished procedures in our laboratory and will describe the application of affinity purification of presynaptic proteins via GST-fusion protein pull-downs for the study of presynaptic phospho-signalling. The pull-down is followed by on-bead proteolysis, chemical stable isotope labelling for precise quantification, enrichment of phosphorylated peptides using TiO2, and LC-MS/MS. The method outlines the software used for peptide/protein identification and quantification, as well as some of the bioinformatics tools that are available for putting the regulated phosphorylation sites into a functional context.

U2 - 10.1007/978-1-4939-8739-9_10

DO - 10.1007/978-1-4939-8739-9_10

M3 - Book chapter

SN - 978-1-4939-9379-6

T3 - Neuromethods

SP - 165

EP - 191

BT - Synaptosomes

A2 - Murphy, Kathryn M.

PB - Springer

ER -

ID: 283984775