Quantitative modeling of self-oligomerization of proteins in the nuclear envelope by fluorescence fluctuation analysis.
Journal
Analytical biochemistry
ISSN: 1096-0309
Titre abrégé: Anal Biochem
Pays: United States
ID NLM: 0370535
Informations de publication
Date de publication:
01 10 2019
01 10 2019
Historique:
received:
19
05
2019
revised:
02
07
2019
accepted:
03
07
2019
pubmed:
8
7
2019
medline:
23
7
2020
entrez:
8
7
2019
Statut:
ppublish
Résumé
Analysis of fluorescence fluctuation data through the time-shifted mean-segmented Q (tsMSQ) analysis method has recently been shown to successfully identify protein oligomerization and mobility in the nuclear envelope by properly accounting for local volume fluctuations of the nuclear envelope within living cells. However, by its nature, tsMSQ produces correlated data which poses unique challenges for applying goodness of fit tests and obtaining parameter uncertainties from individual measurements. In this paper, we overcome these challenges by introducing bootstrap tsMSQ which involves randomly resampling the fluorescence intensity data to eliminate the correlations in the tsMSQ data. This analysis technique was verified in both the cytoplasm and the lumen of the nuclear envelope with well-characterized proteins that served as model systems. Uncertainties and goodness of fit tests of individual measurements were compared to estimates obtained from sampling multiple experiments. We further applied bootstrapping to fluorescence fluctuation data of the luminal domain of the SUN domain-containing protein 2 in order to characterize its self-oligomerization within the nuclear envelope. Analysis of the concentration-dependent brightness suggests a monomer-trimer transition of the protein.
Identifiants
pubmed: 31279795
pii: S0003-2697(19)30473-7
doi: 10.1016/j.ab.2019.113359
pmc: PMC6679984
mid: NIHMS1534675
pii:
doi:
Substances chimiques
Intracellular Signaling Peptides and Proteins
0
Membrane Proteins
0
Nuclear Proteins
0
SUN2 protein, human
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
113359Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM064589
Pays : United States
Informations de copyright
Copyright © 2019 Elsevier Inc. All rights reserved.
Références
Biophys J. 2002 Jan;82(1 Pt 1):133-44
pubmed: 11751302
J Biophys Biochem Cytol. 1955 May 25;1(3):257-70
pubmed: 13242591
Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15492-7
pubmed: 14673112
J Biol Chem. 2004 May 14;279(20):21302-11
pubmed: 14764589
Biophys J. 2005 Nov;89(5):3531-47
pubmed: 16143637
J Cell Biol. 2006 Jan 2;172(1):41-53
pubmed: 16380439
Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3147-52
pubmed: 17307882
Opt Lett. 1979 Jul 1;4(7):205-7
pubmed: 19687850
J Cell Sci. 2010 Jun 15;123(Pt 12):1973-8
pubmed: 20519579
Biophys J. 2010 Aug 4;99(3):979-88
pubmed: 20682277
Mol Biol Cell. 2010 Dec;21(24):4306-11
pubmed: 21160072
J Biol Chem. 2012 Feb 17;287(8):5317-26
pubmed: 22170055
Cell. 2012 May 25;149(5):1035-47
pubmed: 22632968
Biophys J. 2014 Jul 1;107(1):66-75
pubmed: 24988342
Prog Mol Biol Transl Sci. 2014;126:205-15
pubmed: 25081619
Anal Biochem. 2015 Jul 1;480:11-20
pubmed: 25862080
Nat Commun. 2015 Jun 16;6:7314
pubmed: 26076669
PLoS One. 2015 Jun 22;10(6):e0130063
pubmed: 26099032
Structure. 2016 Jan 5;24(1):80-91
pubmed: 26688217
J Cell Biol. 2016 Nov 21;215(4):575-590
pubmed: 27831485
Biophys J. 2017 Jul 11;113(1):138-147
pubmed: 28700912
J Neural Transm (Vienna). 2018 Feb;125(2):177-191
pubmed: 29196815
Mol Biol Cell. 2018 May 1;29(9):1003-1011
pubmed: 29514929
Methods Mol Biol. 2018;1840:121-135
pubmed: 30141043
Methods. 2019 Mar 15;157:28-41
pubmed: 30268407
Biochemistry. 1996 Nov 19;35(46):14480-5
pubmed: 8931544