Methods for characterizing protein acetylation during viral infection.
Acetylome
HDAC
Mass spectrometry
PRM
Proteomics
SIRT
Sirtuin
Targeted mass spectrometry
Journal
Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271
Informations de publication
Date de publication:
2019
2019
Historique:
entrez:
14
10
2019
pubmed:
14
10
2019
medline:
3
6
2020
Statut:
ppublish
Résumé
Lysine acetylation is a prevalent posttranslational modification that acts as a regulator of protein function, subcellular localization, and interactions. A growing body of work has highlighted the importance of temporal alterations in protein acetylation during infection with a range of human viruses. It has become clear that both cellular and viral proteins are decorated by lysine acetylations, and that these modifications contribute to core host defense and virus replication processes. Further defining the extent and dynamics of protein acetylation events during the progression of an infection can provide an important new perspective on the intricate mechanisms underlying the biology and pathogenesis of virus infections. Here, we provide protocols for identifying, quantifying, and probing the regulation of lysine acetylations during viral infection. We describe the use of acetyl-lysine immunoaffinity purification and quantitative mass spectrometry for assessing the cellular acetylome at different stages of an infection. As an alternative to traditional antibody-mediated western blotting, we discuss the benefits of targeted mass spectrometry approaches for detecting and quantifying site-specific acetylations on proteins of interest. Specifically, we provide a protocol using parallel reaction monitoring (PRM). We further discuss experimental considerations that are specific to studying viral infections. Finally, we provide a brief overview of the types of assays that can be employed to characterize the function of an acetylation event in the context of infection. As a method to interrogate the regulation of acetylation, we describe the Fluor de Lys assay for monitoring the enzymatic activities of deacetylases.
Identifiants
pubmed: 31606092
pii: S0076-6879(19)30280-0
doi: 10.1016/bs.mie.2019.06.030
pmc: PMC6889803
mid: NIHMS1059905
pii:
doi:
Substances chimiques
Proteins
0
Proteome
0
Viral Proteins
0
Lysine
K3Z4F929H6
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
587-620Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM114141
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007388
Pays : United States
Informations de copyright
© 2019 Elsevier Inc. All rights reserved.
Références
Nat Rev Mol Cell Biol. 2014 Aug;15(8):536-50
pubmed: 25053359
Nat Commun. 2018 Nov 23;9(1):4967
pubmed: 30470744
EMBO J. 2002 Jun 3;21(11):2715-23
pubmed: 12032084
Trends Microbiol. 2013 Jun;21(6):277-85
pubmed: 23517573
J Virol. 2017 Jul 27;91(16):
pubmed: 28615197
Int J Mol Sci. 2018 Aug 04;19(8):null
pubmed: 30081552
J Immunol. 2006 Dec 15;177(12):8658-66
pubmed: 17142766
Viruses. 2013 Jun 27;5(7):1607-32
pubmed: 23807710
Cell Cycle. 2011 Nov 1;10(21):3701-5
pubmed: 22033337
Oncogene. 2011 Feb 17;30(7):865-75
pubmed: 20935676
J Virol. 2017 Aug 24;91(18):
pubmed: 28679763
J Biol Chem. 2013 Oct 4;288(40):29036-45
pubmed: 23946487
J Virol. 2008 Oct;82(19):9525-36
pubmed: 18653451
Proc Natl Acad Sci U S A. 2001 Sep 11;98(19):10572-7
pubmed: 11535832
Mol Syst Biol. 2014 Jan 30;10:716
pubmed: 24489116
PLoS One. 2013 Jun 27;8(6):e67186
pubmed: 23826228
J Biol Chem. 2003 Apr 4;278(14):12310-8
pubmed: 12501250
Nat Commun. 2015 Mar 13;6:6537
pubmed: 26055519
Mol Syst Biol. 2008;4:222
pubmed: 18854821
J Virol. 2011 Aug;85(15):7474-82
pubmed: 21450817
Methods Mol Biol. 2016;1436:47-84
pubmed: 27246208
Proc Natl Acad Sci U S A. 2019 Feb 26;116(9):3752-3757
pubmed: 30808761
J Virol. 2006 Apr;80(7):3189-204
pubmed: 16537587
J Proteomics. 2014 Apr 4;100:147-59
pubmed: 24200835
EMBO J. 2004 Jun 2;23(11):2269-80
pubmed: 15141169
PLoS One. 2015 Jun 22;10(6):e0131169
pubmed: 26098117
Science. 2009 Aug 14;325(5942):834-40
pubmed: 19608861
Cancer Res. 2004 Feb 1;64(3):1079-86
pubmed: 14871841
Nat Commun. 2017 Nov 2;8(1):1259
pubmed: 29097654
Cell. 2014 Dec 18;159(7):1615-25
pubmed: 25525879
Proc Natl Acad Sci U S A. 2012 Jun 26;109(26):10558-63
pubmed: 22691496
Mol Cell Proteomics. 2014 Jan;13(1):168-83
pubmed: 24163442
Nat Protoc. 2007;2(8):1896-906
pubmed: 17703201
Virology. 2000 Nov 25;277(2):278-95
pubmed: 11080476
mSystems. 2016 May;1(3):null
pubmed: 27376153
Sci Rep. 2016 Dec 22;6:39787
pubmed: 28004786
Mol Cell. 2006 Aug;23(4):607-18
pubmed: 16916647
J Biol Chem. 2002 Jun 21;277(25):22215-21
pubmed: 11956210
Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9280-5
pubmed: 12883000
Mol Cell Proteomics. 2012 Oct;11(10):1048-62
pubmed: 22826441
Mol Syst Biol. 2013;9:672
pubmed: 23752268
MBio. 2014 Dec 16;5(6):null
pubmed: 25516616
Oncotarget. 2016 Jan 12;7(2):1687-706
pubmed: 26625199
J Virol. 2007 Dec;81(23):12740-7
pubmed: 17898049
Curr Biol. 1999 Dec 16-30;9(24):1489-92
pubmed: 10607594
Mol Cell Proteomics. 2012 Nov;11(11):1475-88
pubmed: 22865924
Mol Cell Proteomics. 2013 Nov;12(11):3237-52
pubmed: 23938468
Dev Cell. 2018 Apr 9;45(1):83-100.e7
pubmed: 29634939
EMBO J. 1999 Nov 1;18(21):6106-18
pubmed: 10545121
Nat Rev Mol Cell Biol. 2004 May;5(5):392-401
pubmed: 15122352
J Virol. 2014 Oct;88(19):11229-39
pubmed: 25031336
J Biol Chem. 2015 Jan 23;290(4):2368-78
pubmed: 25451941
PLoS One. 2014 Jul 15;9(7):e101762
pubmed: 25025332
EMBO J. 2016 Feb 15;35(4):429-42
pubmed: 26746851
Anal Chem. 2003 Feb 1;75(3):663-70
pubmed: 12585499
Mol Cell Proteomics. 2015 Sep;14(9):2429-40
pubmed: 25953088