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Quantification of endogenous and exogenous protein expressions of Na,K-ATPase with super-resolution PALM/STORM imaging
KTH, School of Engineering Sciences (SCI), Applied Physics, Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Engineering Sciences (SCI), Applied Physics, Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-5584-9170
KTH, School of Engineering Sciences (SCI), Applied Physics, Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab. 2 Science for Life Laboratory, Department of Women’s and Children’s Health, Karolinska Institutet, Stockholm, Sweden.ORCID iD: 0000-0003-0578-4003
2018 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203Article in journal, Letter (Refereed) Published
Abstract [en]

Transient transfection of fluorescent fusion proteins is a key enabling technology in fluorescent microscopy to spatio-temporally map cellular protein distributions. Transient transfection of proteins may however bypass normal regulation of expression, leading to overexpression artefacts like misallocations and excess amounts. In this study we investigate the use of STORM and PALM microscopy to quantitatively monitor endogenous and exogenous protein expression. Through incorporation of an N-terminal hemagglutinin epitope to a mMaple3 fused Na,K-ATPase (α1 isoform), we analyze the spatial and quantitative changes of plasma membrane Na,K-ATPase localization during competitive transient expression. Quantification of plasma membrane protein density revealed a time dependent increase of Na,K-ATPase, but no increase in size of protein clusters. Results show that after 41h transfection, the total plasma membrane density of Na,K-ATPase increased by 63% while the endogenous contribution was reduced by 16%.

Place, publisher, year, edition, pages
2018.
National Category
Nano Technology
Research subject
Biological Physics
Identifiers
URN: urn:nbn:se:kth:diva-258007DOI: 10.1371/journal. pone.0195825OAI: oai:DiVA.org:kth-258007DiVA, id: diva2:1349647
Funder
Swedish Research Council, 2013-6041Swedish Research Council, 2015-4198Swedish Foundation for Strategic Research , RIF14-0091
Note

QC 20190916

Available from: 2019-09-09 Created: 2019-09-09 Last updated: 2019-09-16Bibliographically approved

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Bernhem, KristofferBlom, HansBrismar, Hjalmar

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