Marine Biotechnology

, Volume 13, Issue 3, pp 496–504

Shotgun Proteomic Analysis of Emiliania huxleyi, a Marine Phytoplankton Species of Major Biogeochemical Importance

  • Bethan M. Jones
  • Richard J. Edwards
  • Paul J. Skipp
  • C. David O’Connor
  • M. Debora Iglesias-Rodriguez
Original Article

DOI: 10.1007/s10126-010-9320-0

Cite this article as:
Jones, B.M., Edwards, R.J., Skipp, P.J. et al. Mar Biotechnol (2011) 13: 496. doi:10.1007/s10126-010-9320-0

Abstract

Emiliania huxleyi is a unicellular marine phytoplankton species known to play a significant role in global biogeochemistry. Through the dual roles of photosynthesis and production of calcium carbonate (calcification), carbon is transferred from the atmosphere to ocean sediments. Almost nothing is known about the molecular mechanisms that control calcification, a process that is tightly regulated within the cell. To initiate proteomic studies on this important and phylogenetically remote organism, we have devised efficient protein extraction protocols and developed a bioinformatics pipeline that allows the statistically robust assignment of proteins from MS/MS data using preexisting EST sequences. The bioinformatics tool, termed BUDAPEST (Bioinformatics Utility for Data Analysis of Proteomics using ESTs), is fully automated and was used to search against data generated from three strains. BUDAPEST increased the number of identifications over standard protein database searches from 37 to 99 proteins when data were amalgamated. Proteins involved in diverse cellular processes were uncovered. For example, experimental evidence was obtained for a novel type I polyketide synthase and for various photosystem components. The proteomic and bioinformatic approaches developed in this study are of wider applicability, particularly to the oceanographic community where genomic sequence data for species of interest are currently scarce.

Keywords

BUDAPESTEmiliania huxleyiEST analysisMass spectrometryShotgun proteomics

Supplementary material

10126_2010_9320_MOESM1_ESM.doc (60 kb)
ESM Supplementary Table 1Protein identifications from taxonomically restricted protein database (DOC 60.5 kb)
10126_2010_9320_MOESM2_ESM.doc (81 kb)
ESM Supplementary Table 2Consensus protein identifications from BUDAPEST pipeline (DOC 103 kb)
10126_2010_9320_MOESM3_ESM.doc (31 kb)
ESM Supplementary resultsSupplementary results detailing 1) the process and 2) location proteins identified for E. huxleyi strains NZEH, CCMP371 and CCMP1516 with respect to available GO terms (DOC 31 kb)
10126_2010_9320_MOESM4_ESM.zip (1.1 mb)
ESM data 1Trees and alignments generated from taxonomically restricted protein database search dataset (ZIP 1.05 mb)
10126_2010_9320_MOESM5_ESM.zip (4 mb)
ESM data 2Sequences, trees and alignments generated from BUDAPEST analysis (ZIP 4 mb)
10126_2010_9320_MOESM6_ESM.xls (1.7 mb)
ESM data 3Excel worksheets showing raw Mascot results for the searches of MS/MS data against the Emiliania huxley ESTs and taxonomically restricted protein datasets. Also included are peptide, BUDAPEST and taxonomically restricted database worksheets and results (XLS 1.72 mb)

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Bethan M. Jones
    • 1
  • Richard J. Edwards
    • 2
  • Paul J. Skipp
    • 2
    • 3
  • C. David O’Connor
    • 2
    • 3
  • M. Debora Iglesias-Rodriguez
    • 1
  1. 1.School of Ocean and Earth Science, National Oceanography Centre, SouthamptonUniversity of SouthamptonSouthamptonUK
  2. 2.School of Biological SciencesUniversity of SouthamptonSouthamptonUK
  3. 3.Centre for Proteomic ResearchUniversity of SouthamptonSouthamptonUK