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Biologia Plantarum

, Volume 55, Issue 3, pp 499–506 | Cite as

Impact of homogenization and protein extraction conditions on the obtained tobacco pollen proteomic patterns

  • J. Fíla
  • V. Čapková
  • J. Feciková
  • D. HonysEmail author
Original Papers

Abstract

Mature pollen grain represents a highly desiccated structure with an extremely tough cell wall. Thanks to it, it resists common proteomic protocols. Instead, a robust homogenization has to be performed since proteins are needed to burst out of the cell to be included in the extracted proteome fraction. Here, a novel way of pollen homogenization employing Roche MagNA Lyser Instrument is presented, sparing time and laborious work. However, plant proteomics does not rely solely on perfect homogenization; also the choice of the extraction protocol is of key importance. The composition of the extraction buffer has a decisive influence on which proteome fraction will be extracted. Therefore the second part of our study is dedicated to the comparison of different extraction protocols with respect to subsequent proteomic analyses.

Additional key words

proteomics Roche MagNA Lyser Instrument 

Abbreviations

BSA

bovine serum albumine

CHAPS

3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate

DTT

dithiothreitol

IEF

isoelectric focusing

PMSF

phenylmethanesulphonyl fluoride

SB 3-12

sulfobetaine-12

SDS-PAGE

sodium-dodecyl-sulfate polyacrylamide gel electrophoresis

TCA

trichloroacetic acid

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Notes

Acknowledgements

This work was supported by ESF (COST Action FA 0603), the Ministry of Education of the Czech Republic (MSMT grant No. OC08011) and Czech Science Foundation (grant No. P501/11/1462).

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • J. Fíla
    • 1
    • 2
  • V. Čapková
    • 1
  • J. Feciková
    • 1
  • D. Honys
    • 1
    Email author
  1. 1.Laboratory of Pollen Biology, Institute of Experimental BotanyAcademy of Sciences of the Czech RepublicPragueCzech Republic
  2. 2.Department of Plant Experimental Biology, Faculty of ScienceCharles University in PraguePragueCzech Republic

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