| RRC ID |
68742
|
| 著者 |
Oura M, Yamamoto J, Ishikawa H, Mikuni S, Fukushima R, Kinjo M.
|
| タイトル |
Polarization-dependent fluorescence correlation spectroscopy for studying structural properties of proteins in living cell.
|
| ジャーナル |
Sci Rep
|
| Abstract |
Rotational diffusion measurement is predicted as an important method in cell biology because the rotational properties directly reflect molecular interactions and environment in the cell. To prove this concept, polarization-dependent fluorescence correlation spectroscopy (pol-FCS) measurements of purified fluorescent proteins were conducted in viscous solution. With the comparison between the translational and rotational diffusion coefficients obtained from pol-FCS measurements, the hydrodynamic radius of an enhanced green fluorescent protein (EGFP) was estimated as a control measurement. The orientation of oligomer EGFP in living cells was also estimated by pol-FCS and compared with Monte Carlo simulations. The results of this pol-FCS experiment indicate that this method allows an estimation of the molecular orientation using the characteristics of rotational diffusion. Further, it can be applied to analyze the degree of molecular orientation and multimerization or detection of tiny aggregation of aggregate-prone proteins.
|
| 巻・号 |
6
|
| ページ |
31091
|
| 公開日 |
2016-8-4
|
| DOI |
10.1038/srep31091
|
| PII |
srep31091
|
| PMID |
27489044
|
| PMC |
PMC4973283
|
| MeSH |
Animals
COS Cells
Chlorocebus aethiops
Fluorescence Polarization
Green Fluorescent Proteins / chemistry*
Hydrodynamics
Molecular Dynamics Simulation
Monte Carlo Method
Protein Aggregates
Protein Structure, Quaternary
Recombinant Proteins / chemistry
Spectrometry, Fluorescence / methods*
|
| IF |
3.998
|
| リソース情報 |
| 遺伝子材料 |
FL-linked EGFP dimer (RDB19236)
FL-linked EGFP trimer (RDB19237)
FL-linked EGFP tetramer (RDB19238)
FL-linked EGFP pentamer (RDB19239) |
