Plant Science


    Protocols in Current Issue
    In vitro Auto- and Substrate-Ubiquitination Assays
    Authors:  Hye Lin Park, Han Yong Lee and Gyeong Mee Yoon, date: 04/05/2022, view: 986, Q&A: 1

    The precise regulation of the homeostasis of the cellular proteome is critical for the appropriate growth and development of plants. It also allows the plants to respond to various environmental stresses, by modulating their biochemical and physiological aspects in a timely manner. Ubiquitination of cellular proteins is one of the major

    Simple Method to Determine Protein Redox State in Arabidopsis thaliana
    Authors:  Keisuke Yoshida and Toru Hisabori, date: 06/05/2019, view: 5591, Q&A: 1
    [Abstract] Thiol-based redox regulation is a posttranslational protein modification that plays a key role in many biological aspects. To understand its regulatory functions, we need a method to directly assess protein redox state in vivo. Here we present a simple procedure to determine protein redox state in a model plant Arabidopsis thaliana ...
    Detection of Disulfides in Protein Extracts of Arabidopsis thaliana Using Monobromobimane (mBB)
    Authors:  Shin‐nosuke Hashida and Maki Kawai-Yamada, date: 03/05/2019, view: 5034, Q&A: 0
    [Abstract] Thiol-disulfide exchange is a key posttranslational modification, determining the folding process of intra- and inter-protein structures. Thiols can be detected by colorimetric reagents, which are stoichiometrically reduced by free thiols, and by fluorescent adducts, showing fluorescence only after thioester formation. We adapted a simple ...
    Histone Deubiquitination Assay in Nicotiana benthamiana
    Authors:  Shujing Liu and Lars Hennig, date: 03/05/2018, view: 5048, Q&A: 0
    [Abstract] Histone modifications are a group of post-translational modifications on histones which can alter chromatin structure and affect gene expression. Histone ubiquitination is a histone modification found in particular on histone H2A and H2B. Histone ubiquitination can be reversed by ubiquitin-specific proteases (UBP). Here, we describe an in vivo ...
    In vitro Ubiquitin Dimer Formation Assay
    Authors:  Sheng Wang, Ling Cao and Hong Wang, date: 01/05/2017, view: 7318, Q&A: 0
    [Abstract] The process of protein ubiquitination typically consists of three sequential steps to add an ubiquitin (Ub) or Ub chain to a substrate protein, requiring three different enzymes, ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), and ubiquitin protein ligase (E3). Most E2s possess the classical E2 activity in forming E2-Ub ...
    A Phosphopeptide Purification Protocol for the Moss Physcomitrella paten
    Authors:  Xiaoqin Wang and Yikun He, date: 07/20/2015, view: 8636, Q&A: 0
    [Abstract] Protein phosphorylation is one of the most common post-translational modifications in eukaryotic cells and plays a critical role in a vast array of cellular processes. Efficient methods of protein extraction and phosphopeptide purification are required to ensure the detection of high quality of proteins. In our hands, phenol extraction of proteins ...
    In vitro Detection of S-acylation on Recombinant Proteins via the Biotin-Switch Technique
    Authors:  Dong Qi and Roger W. Innes, date: 11/20/2014, view: 8751, Q&A: 0
    [Abstract] Protein palmitoylation is the post-translational modification of proteins via the attachment of palmitate through acyl linkages. The nucleophile sulfhydryl group of cysteines is the common palmitoylation site. Covalent attachment of palmitate occurs on numerous proteins and is usually associated with directing protein localization to the ...
    In vitro Protein Ubiquitination Assays
    Authors:  Qingzhen Zhao and Qi Xie, date: 10/05/2013, view: 44203, Q&A: 1
    [Abstract] Ubiquitin can be added to substrate protein as a protein tag by the concerted actions of ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2) and ubiquitin protein ligase (E3). At the present of E1 and ubiquitin, E2 activity can be determined by the thio-ester formation. The E3 activity of a putative protein as well as the E2/E3 or ...

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