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Protocol for evaluating protein relocalization from the plasma membrane to chloroplasts
- Source :
- STAR Protocols, STAR Protocols, Vol 2, Iss 4, Pp 100816-(2021)
- Publication Year :
- 2021
-
Abstract
- Summary We present a protocol for analyzing and evaluating the relocalization of proteins from the plasma membrane to chloroplasts. Some plant membrane-bound proteins carry dual targeting signals, e.g., a membrane-anchoring N-myristoylation motif and a chloroplast transit peptide. These proteins are predominantly targeted to membranes; upon certain cues, however, they can undergo detachment from membranes and relocalization to chloroplasts. This protocol combines imaging and biochemical analyses to track in a reliable and quantitative manner the relocalization of proteins between subcellular organelles. For complete details on the use and execution of this protocol, please refer to Medina-Puche et al. (2020).<br />Graphical abstract<br />Highlights • A method for tracking relocation of proteins from plasma membrane-to-chloroplasts • A method for a quantitative evaluation of the localization in a dynamic manner • Relocation of both host and foreign proteins can be studied • The protocol integrates biochemical and imaging approaches<br />We present a protocol for analyzing and evaluating the relocalization of proteins from the plasma membrane to chloroplasts. Some plant membrane-bound proteins carry dual targeting signals, e.g., a membrane-anchoring N-myristoylation motif and a chloroplast transit peptide. These proteins are predominantly targeted to membranes; upon certain cues, however, they can undergo detachment from membranes and relocalization to chloroplasts. This protocol combines imaging and biochemical analyses to track in a reliable and quantitative manner the relocalization of proteins between subcellular organelles.
- Subjects :
- Science (General)
Chloroplasts
Dual targeting
General Biochemistry, Genetics and Molecular Biology
Cell membrane
Q1-390
Chloroplast Proteins
Transit Peptide
Protein Biochemistry
Organelle
Tobacco
medicine
Image Processing, Computer-Assisted
Protocol
Plant Proteins
Microscopy
General Immunology and Microbiology
Chemistry
General Neuroscience
Cell Membrane
food and beverages
Membrane Proteins
Cell Biology
Cell biology
Molecular Imaging
Chloroplast
Membrane
medicine.anatomical_structure
Cell separation/fractionation
Plant sciences
Subjects
Details
- ISSN :
- 26661667
- Volume :
- 2
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- STAR protocols
- Accession number :
- edsair.doi.dedup.....da7bcc0949bb549208f5a3bf79509528