Biochemistry (Moscow)

, Volume 74, Issue 5, pp 480–490 | Cite as

Proteome analysis of the moss Physcomitrella patens (Hedw.) B.S.G.

  • A. Yu. SkripnikovEmail author
  • N. B. Polyakov
  • E. V. Tolcheva
  • V. V. Velikodvorskaya
  • S. V. Dolgov
  • I. A. Demina
  • M. A. Rogova
  • V. M. Govorun


The sequencing of the moss Physcomitrella patens genome has facilitated studies of the plant proteome. To develop a proteome reference map based on the genome sequence, we conducted 2D electrophoreses of proteins extracted from moss protoplasts, protonemata, and gametophores grown under standard conditions on Petri dishes. On silver-stained gels, depending on the developmental stage of the moss, we resolved from 500 to 600 protein spots that were then excised and digested by trypsin, and 212 proteins were identified by PMF-MALDI-TOF. To enhance the proteome coverage, we performed 1D SDS-PAGE with subsequent separation of tryptic peptides derived from digested gel band slices by LC-ESI-MS/MS. The proposed approach allowed us to identify 186 proteins had not been determined by 2D PMF-MALDI-TOF. Proteins identified by both methods were categorized using a system of clusterization of orthologous genes as metabolism (26%), cellular processes and signaling (16%), and information storage and processing (7%). Proteome analysis by differential gel electrophoresis revealed moderate differences between filamentous protonemata and leafy shoots. Surprisingly, protoplasts isolated from protonema filaments displayed significant differences in protein composition compared with both protonemata and gametophores.

Key words

proteomics 2D electrophoresis MALDI-TOF LC-ESI-MS/MS Physcomitrella patens protoplasts DiGE 



differential gel electrophoresis




EuKaryotic Orthologous Groups


liquid chromatography-electrospray ionization-tandem mass spectrometry


matrix-assisted laser desorption/ionization time-of-flight mass spectrometry


polyacrylamide gel electrophoresis


peptide mass fingerprinting


ribulose bisphosphate carboxylase


sodium dodecyl sulfate


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Supplementary material

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Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • A. Yu. Skripnikov
    • 1
    • 2
    Email author
  • N. B. Polyakov
    • 1
  • E. V. Tolcheva
    • 1
  • V. V. Velikodvorskaya
    • 1
  • S. V. Dolgov
    • 3
  • I. A. Demina
    • 4
  • M. A. Rogova
    • 4
  • V. M. Govorun
    • 1
    • 4
  1. 1.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Biology FacultyLomonosov Moscow State UniversityMoscowRussia
  3. 3.Branch of Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesPushchino, Moscow RegionRussia
  4. 4.Scientific Research Institute of Physicochemical MedicineMoscowRussia

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