An affinity pull-down approach to identify the plant cyclic nucleotide interactome
Permanent link to this recordhttp://hdl.handle.net/10754/562961
MetadataShow full item record
AbstractCyclic nucleotides (CNs) are intracellular second messengers that play an important role in mediating physiological responses to environmental and developmental signals, in species ranging from bacteria to humans. In response to these signals, CNs are synthesized by nucleotidyl cyclases and then act by binding to and altering the activity of downstream target proteins known as cyclic nucleotide-binding proteins (CNBPs). A number of CNBPs have been identified across kingdoms including transcription factors, protein kinases, phosphodiesterases, and channels, all of which harbor conserved CN-binding domains. In plants however, few CNBPs have been identified as homology searches fail to return plant sequences with significant matches to known CNBPs. Recently, affinity pull-down techniques have been successfully used to identify CNBPs in animals and have provided new insights into CN signaling. The application of these techniques to plants has not yet been extensively explored and offers an alternative approach toward the unbiased discovery of novel CNBP candidates in plants. Here, an affinity pull-down technique for the identification of the plant CN interactome is presented. In summary, the method involves an extraction of plant proteins which is incubated with a CN-bait, followed by a series of increasingly stringent elutions that eliminates proteins in a sequential manner according to their affinity to the bait. The eluted and bait-bound proteins are separated by one-dimensional gel electrophoresis, excised, and digested with trypsin after which the resultant peptides are identified by mass spectrometry - techniques that are commonplace in proteomics experiments. The discovery of plant CNBPs promises to provide valuable insight into the mechanism of CN signal transduction in plants. © Springer Science+Business Media New York 2013.
JournalMethods in Molecular Biology
- The arabidopsis cyclic nucleotide interactome.
- Authors: Donaldson L, Meier S, Gehring C
- Issue date: 2016 May 11
- Cyclic nucleotides.
- Authors: Newton RP, Smith CJ
- Issue date: 2004 Sep
- Protein complexes characterization in Arabidopsis thaliana by tandem affinity purification coupled to mass spectrometry analysis.
- Authors: Bigeard J, Pflieger D, Colcombet J, Gérard L, Mireau H, Hirt H
- Issue date: 2014
- Detection of reactive oxygen species downstream of cyclic nucleotide signals in plants.
- Authors: Walker RK, Berkowitz GA
- Issue date: 2013
- Cyclic nucleotides as affinity tools: phosphorothioate cAMP analogues address specific PKA subproteomes.
- Authors: Hanke SE, Bertinetti D, Badel A, Schweinsberg S, Genieser HG, Herberg FW
- Issue date: 2011 Jul
Showing items related by title, author, creator and subject.
Ionic H-bonding organocatalysts for the ring-opening polymerization of cyclic esters and cyclic carbonatesXu, Jiaxi; Wang, Xin; Liu, Jingjing; Feng, Xiaoshuang; Gnanou, Yves; Hadjichristidis, Nikos (Progress in Polymer Science, Elsevier BV, 2021-11-10) [Article]Ring-opening polymerization (ROP) of cyclic monomers is a prevalent and convenient method for the synthesis of well-defined polymers with initiators/catalysts that promote a nucleophilic or electrophilic attack on the monomers. Selective activation of functional groups or linkages of the monomer without those carried out in the polymer chains, especially at high conversion, is one of the challenges faced by ROP catalysts. H-bonding organocatalysts can offer precise selectivity for ROP in a wide range of monomers. The firstly reported neutral H-bonding organocatalysts are characterized by high selectivity but long reaction time and low reactivity. In contrast, ionic H-bonding organocatalysts, which have extensively developed over the last ten years, exhibit fast polymerization rates and high selectivity. Besides, some ionic H-bonding organocatalysts with good thermal stability and high reactivity can be used in a wide range of ROP temperatures (-60 °C to over 200 °C). Furthermore, ionic H-bonding organocatalysts comply with biosafety principles promoted by green chemistry. This review covers the mechanistic insights (monomer activation, initiator/chain-end activation, synergistic activation, and bifunctional activation) of ionic H-bonding organocatalytic ROP, as well as the strategies for monomer and initiator/chain-end activation.
A "catalyst switch" Strategy for the sequential metal-free polymerization of epoxides and cyclic Esters/CarbonateZhao, Junpeng; Pahovnik, David; Gnanou, Yves; Hadjichristidis, Nikos (Macromolecules, American Chemical Society (ACS), 2014-06-03) [Article]A "catalyst switch" strategy was used to synthesize well-defined polyether-polyester/polycarbonate block copolymers. Epoxides (ethylene oxide and/or 1,2-butylene oxide) were first polymerized from a monoalcohol in the presence of a strong phosphazene base promoter (t-BuP4). Then an excess of diphenyl phosphate (DPP) was introduced, followed by the addition and polymerization of a cyclic ester (ε-caprolactone or δ-valerolactone) or a cyclic carbonate (trimethylene carbonate), where DPP acted as both the neutralizer of phosphazenium alkoxide (polyether chain end) and the activator of cyclic ester/carbonate. This work has provided a one-pot sequential polymerization method for the metal-free synthesis of block copolymers from monomers which are suited for different types of organic catalysts. © 2014 American Chemical Society.
An Efficient and General Strategy toward the Synthesis of Polyethylene-Based Cyclic PolymersJiang, Yu; Zhang, Zhen; Wang, De; Hadjichristidis, Nikos (Macromolecules, American Chemical Society (ACS), 2018-04-13) [Article]A novel strategy toward well-defined polyethylene-based cyclic homo/copolymers is presented. Tris(3-(anthracen-9-ylmethoxy)propyl)borane, prepared by hydroboration of 9-((allyloxy)methyl)anthracene with BH, was used to initiate the polyhomologation of dimethylsulfoxonium methylide to afford well-defined anthracene-teminated linear polyethylene (PE). The azido and alkynyl groups at α and ω positions of the PE chain were introduced via the anthracene/maleimide Diels-Alder (D-A) reaction and esterification, respectively. Subsequent intramolecular