Abstract
Several methodologies have been employed to understand the kinetics of induced autophagic degradation in plants, but most of them are not capable of distinguishing the autophagic cargo proteins before and after induction of autophagy in cells. Here, we designed a mass photoconverter that allowed us to simultaneously monitor protein synthesis and degradation in tobacco BY-2 cells using a photoconvertible fluorescence marker protein, Kikume Green Red (KikGR). An example of a new protocol for the analysis of autophagy progression using a fusion protein of cytochrome b5 and KikGR under phosphate starvation is described. The other example described is the analysis of the proliferation of Golgi apparatus in tobacco BY-2 cells using the fusion protein of a prolyl 4-hydroxylase NtP4H1.1 and monomeric KikGR. A detailed protocol on key analysis, as well as tips and notes for experiments using KikGR proteins, are described.
| Original language | English |
|---|---|
| Title of host publication | Methods in Enzymology |
| Publisher | Academic Press Inc. |
| Pages | 515-526 |
| Number of pages | 12 |
| Volume | 588 |
| DOIs | |
| Publication status | Published - 2017 |
Publication series
| Name | Methods in Enzymology |
|---|---|
| Volume | 588 |
| ISSN (Print) | 0076-6879 |
| ISSN (Electronic) | 1557-7988 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
Cite this
Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants. / Abiodun, M. O.; Matsuoka, Ken.
Methods in Enzymology. Vol. 588 Academic Press Inc., 2017. p. 515-526 (Methods in Enzymology; Vol. 588).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants
AU - Abiodun, M. O.
AU - Matsuoka, Ken
PY - 2017
Y1 - 2017
N2 - Several methodologies have been employed to understand the kinetics of induced autophagic degradation in plants, but most of them are not capable of distinguishing the autophagic cargo proteins before and after induction of autophagy in cells. Here, we designed a mass photoconverter that allowed us to simultaneously monitor protein synthesis and degradation in tobacco BY-2 cells using a photoconvertible fluorescence marker protein, Kikume Green Red (KikGR). An example of a new protocol for the analysis of autophagy progression using a fusion protein of cytochrome b5 and KikGR under phosphate starvation is described. The other example described is the analysis of the proliferation of Golgi apparatus in tobacco BY-2 cells using the fusion protein of a prolyl 4-hydroxylase NtP4H1.1 and monomeric KikGR. A detailed protocol on key analysis, as well as tips and notes for experiments using KikGR proteins, are described.
AB - Several methodologies have been employed to understand the kinetics of induced autophagic degradation in plants, but most of them are not capable of distinguishing the autophagic cargo proteins before and after induction of autophagy in cells. Here, we designed a mass photoconverter that allowed us to simultaneously monitor protein synthesis and degradation in tobacco BY-2 cells using a photoconvertible fluorescence marker protein, Kikume Green Red (KikGR). An example of a new protocol for the analysis of autophagy progression using a fusion protein of cytochrome b5 and KikGR under phosphate starvation is described. The other example described is the analysis of the proliferation of Golgi apparatus in tobacco BY-2 cells using the fusion protein of a prolyl 4-hydroxylase NtP4H1.1 and monomeric KikGR. A detailed protocol on key analysis, as well as tips and notes for experiments using KikGR proteins, are described.
UR - http://www.scopus.com/inward/record.url?scp=85007591241&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85007591241&partnerID=8YFLogxK
U2 - 10.1016/bs.mie.2016.10.040
DO - 10.1016/bs.mie.2016.10.040
M3 - Chapter
C2 - 28237118
AN - SCOPUS:85007591241
VL - 588
T3 - Methods in Enzymology
SP - 515
EP - 526
BT - Methods in Enzymology
PB - Academic Press Inc.
ER -