mEosBrite Are Bright Variants of mEos3.2 Developed by Semirational Protein Engineering

Marathe, P and HS, MS and Nair, D and Bhattacharyya, D (2020) mEosBrite Are Bright Variants of mEos3.2 Developed by Semirational Protein Engineering. In: Journal of Fluorescence, 30 (3). pp. 703-715.

	PDF jou_flu_30-03_703-715_2020.pdf - Published Version Restricted to Registered users only Download (4MB) | Request a copy
	PDF 10895_2020_2537_MOESM1_ESM.pdf - Published Supplemental Material Restricted to Registered users only Download (303kB) | Request a copy
	PDF 10895_2020_2537_MOESM2_ESM.pdf - Published Supplemental Material Restricted to Registered users only Download (73kB) | Request a copy
	PDF 10895_2020_2537_MOESM3_ESM.pdf - Published Supplemental Material Restricted to Registered users only Download (155kB) | Request a copy
	PDF 10895_2020_2537_MOESM4_ESM.pdf - Published Version Restricted to Registered users only Download (98kB) | Request a copy

Abstract

mEos3.2 is a photoconvertible fluorescence protein with comparatively low brightness, which limits its application in live Super resolution microscopy. To address this issue, we have used semi-rational protein engineering to develop mEosBrite, a new class of improved brightness variants. The improvement in the brightness was confirmed by expression in E.coli as well as mammalian cell lines. Furthermore, biophysical characterization suggests that all the three mEosBrite variant proteins display higher quantum yield, truly monomeric form, less cytotoxicity and lower protein aggregation as compared to the wild type mEos3.2 protein. Most importantly, because of their high photoconversion efficiency mEosBrite variants could be an excellent tool for single-molecule and intensity fluctuation based super-resolution microscopy.

Item Type:	Journal Article
Publication:	Journal of Fluorescence
Publisher:	Springer
Additional Information:	The copyright of this article belongs to Springer
Department/Centre:	Division of Biological Sciences > Centre for Neuroscience
Date Deposited:	12 Aug 2020 09:57
Last Modified:	12 Aug 2020 09:57
URI:	http://eprints.iisc.ac.in/id/eprint/65450

Actions (login required)

View Item