0 249

Cited 0 times in

Highly chromophoric dual-terminus labeling of an intrinsically disordered native eukaryotic protein of interest at nanoscale

Authors
 Jung Min Kim  ;  Baik Lin Seong  ;  Jinho Jung 
Citation
 INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, Vol.245 : 125396, 2023-08 
Journal Title
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
ISSN
 0141-8130 
Issue Date
2023-08
MeSH
Escherichia coli / genetics ; Escherichia coli / metabolism ; Eukaryota ; Eukaryotic Cells* ; Fluorescence Resonance Energy Transfer ; Fluorescent Dyes / chemistry ; HeLa Cells ; Humans ; Intrinsically Disordered Proteins* / metabolism
Keywords
Bead assay ; Dual-fluorescent probe ; Eukaryotic protein labeling ; Intrinsically disordered native protein (IDNP). Macromolecular crowding ; Two-color correlation
Abstract
Chemical conjugation of purified proteins of interest (POIs) in Escherichia coli cells is effective for high expression but has limitations for highly chromogenic dual labeling of intrinsically disordered native proteins (IDNPs). Our probes can tag IDNPs using chemical conjugation during protein synthesis and folding while preserving biologically active structures in mammalian cells. We fluorescently labeled IDNPs in mammalian cells using pure fluorescent methionine and ATTO 565-biotin at the N-or C-terminus, respectively. The dual-labeled Tat protein was used as a model for IDNPs in HeLa cells and detected using Ni-NTA beads to estimate its highly chromogenic concentration. We also demonstrated highly chromogenic double labeling of genetically encoded fluorescent-Tat expression in eukaryotic cells using a single fluorescent dye pair with Fo & BULL;rster resonance energy transfer (FRET) ratio and two-color correlation analysis. This study aims to solve native POI processing and achieve ultrasensitive protein folding for biological and ecological applications at the nanoscale.
Full Text
https://www.sciencedirect.com/science/article/pii/S0141813023022900
DOI
10.1016/j.ijbiomac.2023.125396
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Seong, Baik L(성백린)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/198115
사서에게 알리기
  feedback

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse

Links