What does the f-box protein do?
F-box proteins are involved in many plant vegetative and reproduction growth and development. For example, F-box protein-FOA1 involved in abscisic acid (ABA) signaling to affect the seed germination. ACRE189/ACIF1 can regulate cell death and defense when the pathogen is recognized in the Tobacco and Tomato plant.
How many f-box proteins are there?
In mammals, approximately 70 F-box proteins, each able to target multiple substrates, enable SCF complexes to control the levels of many regulatory proteins with diverse functions. F-box protein substrates are recognized through degradation motifs (degrons).
What is the Fbox domain?
The F-Box domain is a 42–48 conserved amino acid domain found at the N-terminus of F-Box proteins. The F-Box domain mediates interaction with SKP1, which links F-Box proteins to the core ubiquitin-ligase complex that is composed of Rbx1, cdc53/Cul1 and the E2 conjugating enzyme cdc34.
What is an F-box gene?
The F-box is a protein motif of approximately 50 amino acids that functions as a site of protein-protein interaction.
What happens after ubiquitination?
The ubiquitin is then transferred to a second enzyme, called ubiquitin-conjugating enzyme (E2). The final transfer of ubiquitin to the target protein is then mediated by a third enzyme, called ubiquitin ligase or E3, which is responsible for the selective recognition of appropriate substrate proteins.
How do you know if your protein is ubiquitinated?
Strong smears or ladders of high molecular species are typically the result of ubiquitination. The degree of ubiquitination of the protein of interest can be assessed by comparing the ratio of ubiquitinated/unmodified target protein in several experiment conditions.
Are misfolded proteins more stable than native ones?
A protein is considered to be misfolded if it cannot achieve its normal native state. The misfolded protein typically contains β-sheets that are organized in a supramolecular arrangement known as a cross-β structure. These β-sheet-rich assemblies are very stable, very insoluble, and generally resistant to proteolysis.
How do you dilute MG132?
Once MG-132 is solubilized, dilute 1 in 100 using sterile culture medium to obtain a solution at 200 µg/ml (420 µM). Do not store dilutions for more than one day.