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Phosphoprotein Enrichment from Tobacco Mature Pollen Crude Protein Extract
J. Fíla, D. Honys,
Jazyk angličtina Země Spojené státy americké
Typ dokumentu časopisecké články
- MeSH
- chromatografie afinitní MeSH
- fosfoproteiny chemie izolace a purifikace metabolismus MeSH
- hydroxid hlinitý chemie MeSH
- posttranslační úpravy proteinů MeSH
- pyl metabolismus MeSH
- rostlinné proteiny chemie izolace a purifikace metabolismus MeSH
- tabák metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Protein phosphorylation was repeatedly shown to be the most dynamic post-translational modification mediated by a huge orchestra of protein kinases and phosphatases. Upon landing on a stigma, pollen grain dehydration and activation are accompanied by changes in protein phosphorylation together with the translation activation of stored mRNAs. To enable studies of the total phosphoproteome, it is usually necessary to apply various enrichment techniques. In this chapter, one of these protocols that worked previously well on tobacco mature pollen is presented in more detail. The method comprises of three basic steps: (1) picking flowers from the flowering tobacco plants (Nicotiana tabacum cv. Samsun), and collection of the shed pollen grains; (2) extraction of total proteins by TCA/acetone; (3) phosphoprotein enrichment by MOAC with aluminum hydroxide matrix. Taken together this protocol describes how to isolate phosphoproteins out of tobacco mature pollen.
Citace poskytuje Crossref.org
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- $a Protein phosphorylation was repeatedly shown to be the most dynamic post-translational modification mediated by a huge orchestra of protein kinases and phosphatases. Upon landing on a stigma, pollen grain dehydration and activation are accompanied by changes in protein phosphorylation together with the translation activation of stored mRNAs. To enable studies of the total phosphoproteome, it is usually necessary to apply various enrichment techniques. In this chapter, one of these protocols that worked previously well on tobacco mature pollen is presented in more detail. The method comprises of three basic steps: (1) picking flowers from the flowering tobacco plants (Nicotiana tabacum cv. Samsun), and collection of the shed pollen grains; (2) extraction of total proteins by TCA/acetone; (3) phosphoprotein enrichment by MOAC with aluminum hydroxide matrix. Taken together this protocol describes how to isolate phosphoproteins out of tobacco mature pollen.
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