Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants

M. O. Abiodun, Ken Matsuoka

Research output: Chapter in Book/Report/Conference proceedingChapter

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 languageEnglish
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages515-526
Number of pages12
Volume588
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Enzymology
Volume588
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

Autophagy
Proteins
Tobacco
Prolyl Hydroxylases
Fusion reactions
Cytochromes b5
Cell Fusion
Golgi Apparatus
Degradation
Starvation
Proteolysis
Fluorescence
Phosphates
Kinetics
Experiments

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Abiodun, M. O., & Matsuoka, K. (2017). Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants. In Methods in Enzymology (Vol. 588, pp. 515-526). (Methods in Enzymology; Vol. 588). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2016.10.040

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 proceedingChapter

Abiodun, MO & Matsuoka, K 2017, Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants. in Methods in Enzymology. vol. 588, Methods in Enzymology, vol. 588, Academic Press Inc., pp. 515-526. https://doi.org/10.1016/bs.mie.2016.10.040
Abiodun MO, Matsuoka K. Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants. In Methods in Enzymology. Vol. 588. Academic Press Inc. 2017. p. 515-526. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2016.10.040
Abiodun, M. O. ; Matsuoka, Ken. / Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants. Methods in Enzymology. Vol. 588 Academic Press Inc., 2017. pp. 515-526 (Methods in Enzymology).
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