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JXB Advance Access published online on June 4, 2004
Journal of Experimental Botany, doi:10.1093/jxb/erh161
© 2004 by Oxford University Press
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Received February 4, 2004; accepted April 5, 2004
© 2004 Society for Experimental Biology

RESEARCH PAPER

Determining protein identity from sieve element sap in Ricinus communis L. by quadrupole time of flight (Q-TOF) mass spectrometry

Alan Barnes 1, Jeffery Bale 1, Chrystala Constantinidou 1, Peter Ashton 1, Anthony Jones 1, and Jeremy Pritchard 1*

1 The University of Birmingham, School of Biosciences, Edgbaston, Birmingham B15 2TT, UK

* To whom correspondence should be addressed. E-mail: J.Pritchard{at}bham.ac.uk.


  Abstract

The phloem transport system is a complex tissue that primarily carries photoassimilate from source to sink. Its function depends on anucleate sieve elements (SE) supported by companion cells (CC). In this study, SE sap was sampled and the protein identity of soluble proteins was determined with the aim of understanding the function of proteins within the conduit. Unlike many plants, SE sap exudes from incisions in the bark of Ricinus communis and, although there is a greater possibility of contamination from tissues other than SE, sap can be obtained in sufficient quantities to separate proteins using 2D electrophoresis. Spots were excised for trypsin digest, then analysed by quadrupole time of flight (Q-TOF) mass spectrometry (MS) and database searched to determine sequence identity. Overall, 18 proteins were identified in the SE-enriched sap. Proteins identified that have not previously been identified directly from SE sap included a glycine-rich RNA-binding protein, metallothionein, phosphoglycerate mutase, and phosphopyruvate hydratase. The potential role of the identified protein in SE function is discussed. The protein identification in this study provides a first step towards the goal of a greater understanding of the function of proteins within the SE.

Key words: Aphid, companion cell, cyclophilin, MS, phloem, protein, quadrupole time-of-flight, QTOF, sieve-element.


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