Characterisation by proteomics of peribacteroid space and peribacteroid membrane preparations from pea (¤Pisum sativum¤) symbiosomes

G. Saalbach, P. Erik, S. Wienkoop

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    The legume Rhizobium symbiosis leads to the formation of a new compartment in the plant cell, the symbiosome. This compartment harbours the bacteroids surrounded by a peribacteroid membrane (PBM) originating from the plant plasma membrane. The PBM and the space between the PBM and the bacteroid membrane, called peribacteroid space (PS), mediate the exchange of metabolites between the symbionts. Proteome analysis was used as an approach to characterise the proteins in the PBM and the PS. A standard differential centrifugation procedure including a Percoll gradient was used for symbiosome isolation from pea root nodules. Proteins in the PBM and PS fractions obtained from the symbiosomes were separated by two-dimensional gel electrophoresis, and 89 spots were analysed by tandem mass spectrometry. The proteins of 46 spots could be identified by database search. The results showed that PS and even PBM preparations from pea symbiosomes always contain abundant amounts of bacteroid proteins as a contaminate. Interestingly, in addition to a few PS/PBM proteins a number of endomembrane proteins (less likely representing a contaminate), including V-ATPase, BIP, and an integral membrane protein known from COPI-coated vesicles, were found in the PBM fraction, supporting the role of the endomembrane system in PBM biogenesis.
    Original languageEnglish
    JournalProteomics
    Volume2
    Issue number3
    Pages (from-to)325-337
    ISSN1615-9853
    DOIs
    Publication statusPublished - 2002

    Fingerprint Dive into the research topics of 'Characterisation by proteomics of peribacteroid space and peribacteroid membrane preparations from pea (¤Pisum sativum¤) symbiosomes'. Together they form a unique fingerprint.

    Cite this