caa a22 4500 445318368 CHVBK 20180317142655.0 cr unu---uuuuu 170323e20110601xx s 000 0 eng 10.1007/s10126-010-9320-0 doi (NATIONALLICENCE)springer-10.1007/s10126-010-9320-0 Shotgun Proteomic Analysis of Emiliania huxleyi , a Marine Phytoplankton Species of Major Biogeochemical Importance [Elektronische Daten] [Bethan Jones, Richard Edwards, Paul Skipp, C. O'Connor, M. Iglesias-Rodriguez] 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. Springer Science+Business Media, LLC, 2010 BUDAPEST nationallicence Emiliania huxleyi nationallicence EST analysis nationallicence Mass spectrometry nationallicence Shotgun proteomics nationallicence Jones Bethan School of Ocean and Earth Science, National Oceanography Centre, Southampton, University of Southampton, Waterfront Campus, European Way, SO14 3ZH, Southampton, UK aut Edwards Richard School of Biological Sciences, University of Southampton, Southampton, UK aut Skipp Paul School of Biological Sciences, University of Southampton, Southampton, UK aut O'Connor C. School of Biological Sciences, University of Southampton, Southampton, UK aut Iglesias-Rodriguez M. School of Ocean and Earth Science, National Oceanography Centre, Southampton, University of Southampton, Waterfront Campus, European Way, SO14 3ZH, Southampton, UK aut Marine Biotechnology Springer-Verlag 13/3(2011-06-01), 496-504 1436-2228 13:3<496 2011 13 10126 https://doi.org/10.1007/s10126-010-9320-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10126-010-9320-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Jones Bethan School of Ocean and Earth Science, National Oceanography Centre, Southampton, University of Southampton, Waterfront Campus, European Way, SO14 3ZH, Southampton, UK aut NATIONALLICENCE 700 1- Edwards Richard School of Biological Sciences, University of Southampton, Southampton, UK aut NATIONALLICENCE 700 1- Skipp Paul School of Biological Sciences, University of Southampton, Southampton, UK aut NATIONALLICENCE 700 1- O'Connor C. School of Biological Sciences, University of Southampton, Southampton, UK aut NATIONALLICENCE 700 1- Iglesias-Rodriguez M. School of Ocean and Earth Science, National Oceanography Centre, Southampton, University of Southampton, Waterfront Campus, European Way, SO14 3ZH, Southampton, UK aut NATIONALLICENCE 773 0- Marine Biotechnology Springer-Verlag 13/3(2011-06-01), 496-504 1436-2228 13:3<496 2011 13 10126 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer