Location: Food and Feed Safety ResearchTitle: Basic characterization of plant actin depolymerizing factors: a simplified, streamlined guide
|SENGUPTA, SONALI - LSU Agcenter|
|Rajasekaran, Kanniah - Rajah|
|BAISAKH, NIRANJAN - LSU Agcenter|
Submitted to: Protocol Exchange
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/21/2019
Publication Date: 11/21/2019
Citation: Sengupta, S., Rajasekaran, K., Baisakh, N. 2019. Basic characterization of plant actin depolymerizing factors: a simplified, streamlined guide. Protocol Exchange. https://doi.org/10.21203/rs.2.16466/v2.
Interpretive Summary: Actin depolymerizing factors (ADFs) are important family of proteins that affects the actin binding protein and their enzymatic activity is essential in important processes including cell division, elongation and movement of cellular components. This manuscript describes how plant ADFs can be characterized in a simplified, streamlined step-by-step protocol to quickly measure the important functions of the ADF proteins in laboratory studies. This information will be useful to other scientists in plant physiology, molecular biology and biotechnology.
Technical Abstract: Actin depolymerizing factors (ADFs) are small monomeric actin-binding proteins that alter the oligomeric state of cellular actin. Members of the ADF family can bind both the G-actin and F-actin in plants, and their functions are regulated by cellular pH, ionic strength and availability of other binding partners. Actin depolymerization activity is reportedly essential for plant viability. By binding to the ADP-bound form of actin, ADFs severe actin filaments and thereby provide more barbed filament ends for polymerization. They also increase the rate of dissociation of F-actin monomer by changing the helical twist of the actin filament. These two activities together make ADF the major regulator of actin dynamics in plant cell. Therefore, it is essential to measure the binding and depolymerization activity of the plant ADFs. Here, we present a simplified, streamlined step-by-step protocol to quickly measure these important functions of the ADF proteins in vitro.