Plant Germline Development

0485921 - ÚEB 2018 RIV DE eng M - Monography Chapter
Fíla, Jan - Honys, David
Phosphoprotein enrichment from tobacco mature pollen crude protein extract.
Plant Germline Development. New York: Humana Press, 2017 - (Schmidt, A.), Roč. 1669 (2017), s. 265-274. ISBN 978-1-4939-7285-2
R&D Projects: GA ČR GA13-06943S; GA ČR(CZ) GA15-16050S
Institutional support: RVO:61389030
Keywords : Aluminum hydroxide * Male gametophyte * Metal-oxide/hydroxide affinity chromatography * moac * Phosphoprotein enrichment * Pollen grain * TCA/acetone protein extraction
OECD category: Plant sciences, botany
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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.
Permanent Link: http://hdl.handle.net/11104/0280839