biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 55:499-506, 2011 | DOI: 10.1007/s10535-011-0116-5

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

J. Fíla1,2, V. Čapková1, J. Feciková1, D. Honys1,*
1 Laboratory of Pollen Biology, Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Prague, Czech Republic
2 Department of Plant Experimental Biology, Faculty of Science, Charles University in Prague, Prague, Czech Republic

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.

Keywords: proteomics; Roche MagNA Lyser Instrument

Received: November 29, 2009; Accepted: April 9, 2010; Published: September 1, 2011  Show citation

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Fíla, J., Čapková, V., Feciková, J., & Honys, D. (2011). Impact of homogenization and protein extraction conditions on the obtained tobacco pollen proteomic patterns. Biologia plantarum, 55(3), 499-506. doi: 10.1007/s10535-011-0116-5
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