A new method for in vivo plant cell imaging with SNAP-tag proteins

August 21, 2020

A team of scientists at the Nagoya University Institute of Transformative Bio-Molecules (WPI-ITbM) have developed a method for visualizing microtubule dynamics and cell membrane protein endocytosis in living plant cells, an important step forward in plant cell biology.

SNAP-tag visualization of in vivo protein dynamics, a method which binds dyes to proteins to allow fluorescent imaging, has made a wide range of contributions to medical and biological study, for example in cancer research. However, live cell imaging using SNAP-tag with synthetic dyes in plant science has been impossible, as synthetic dyes are unable to reach the target proteins due to the presence of the cell wall in plant cells.

In this study, the research team demonstrated that it was possible to perform live cell imaging using SNAP-tag even in plants, using three types of synthetic dyes which bond to the SNAP-tag to mark microtubules, part of the cytoskeleton. They were able to use a particular dye that does not permeate the cell membrane and fluoresces only when it bonds with SNAP-tag to exclusively mark the auxin transporters in the cell membrane, visualizing the process of membrane proteins being taken into the cell (endocytosis) after they had been marked. Thus, they were able to clearly differentiate between transporter proteins which had been newly synthesized inside the cell and those taken into the cell by endocytosis. They then used tobacco cells to find out which of 31 different dyes were able to enter the cell. Interestingly, it was found that 23 out of these 31 were taken into the tobacco cells, that the majority of them could be used with SNAP-tag to mark cytosolic components in plant cells, and that those which could not permeate the cell membrane could be used to mark membrane proteins outside the cell.

This study provides a new technique for the fluorescent marking of proteins in plant cells, and represent an important step forward in plant cell biology research. It is expected to find use in superresolution imaging using extremely light stable markers, and techniques for determining place- and time-specific pH and Ca2+ levels.
-end-


Institute of Transformative Bio-Molecules (ITbM), Nagoya University

Related Proteins Articles from Brightsurf:

New understanding of how proteins operate
A ground-breaking discovery by Centenary Institute scientists has provided new understanding as to the nature of proteins and how they exist and operate in the human body.

Finding a handle to bag the right proteins
A method that lights up tags attached to selected proteins can help to purify the proteins from a mixed protein pool.

Designing vaccines from artificial proteins
EPFL scientists have developed a new computational approach to create artificial proteins, which showed promising results in vivo as functional vaccines.

New method to monitor Alzheimer's proteins
IBS-CINAP research team has reported a new method to identify the aggregation state of amyloid beta (Aβ) proteins in solution.

Composing new proteins with artificial intelligence
Scientists have long studied how to improve proteins or design new ones.

Hero proteins are here to save other proteins
Researchers at the University of Tokyo have discovered a new group of proteins, remarkable for their unusual shape and abilities to protect against protein clumps associated with neurodegenerative diseases in lab experiments.

Designer proteins
David Baker, Professor of Biochemistry at the University of Washington to speak at the AAAS 2020 session, 'Synthetic Biology: Digital Design of Living Systems.' Prof.

Gone fishin' -- for proteins
Casting lines into human cells to snag proteins, a team of Montreal researchers has solved a 20-year-old mystery of cell biology.

Coupled proteins
Researchers from Heidelberg University and Sendai University in Japan used new biotechnological methods to study how human cells react to and further process external signals.

Understanding the power of honey through its proteins
Honey is a culinary staple that can be found in kitchens around the world.

Read More: Proteins News and Proteins Current Events
Brightsurf.com is a participant in the Amazon Services LLC Associates Program, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to Amazon.com.