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  • 1.
    Sialana, Fernando J.
    et al.
    Univ Vienna, Dept Pharmaceut Chem, Vienna, Austria.;Austrian Acad Sci, CeMM Res Ctr Mol Med, Vienna, Austria..
    Gulyassy, Peter
    Eotvos Lorand Univ, Inst Biol, Lab Prote, Budapest, Hungary..
    Majek, Peter
    Austrian Acad Sci, CeMM Res Ctr Mol Med, Vienna, Austria..
    Sjöstedt, Evelina
    KTH, Centres, Science for Life Laboratory, SciLifeLab. Uppsala Univ, Dept Immunol Genet & Pathol, Sci Life Lab, Uppsala, Sweden..
    Kis, Viktor
    Eotvos Lorand Univ, Dept Anat Cell & Dev Biol, Budapest, Hungary..
    Muller, Andre C.
    Austrian Acad Sci, CeMM Res Ctr Mol Med, Vienna, Austria.;ThermoFisher Sci, Vienna, Austria..
    Rudashevskaya, Elena L.
    Med Univ Vienna, Inst Med Chem, Vienna, Austria.;ISAS, Leibniz Inst Analyt Wissensch, Dortmund, Germany..
    Mulder, Jan
    Uppsala Univ, Dept Neurosci, Sci Life Lab, Uppsala, Sweden..
    Bennett, Keiryn L.
    Austrian Acad Sci, CeMM Res Ctr Mol Med, Vienna, Austria..
    Lubec, Gert
    Univ Vienna, Dept Pharmaceut Chem, Vienna, Austria..
    Mass spectrometric analysis of synaptosomal membrane preparations for the determination of brain receptors, transporters and channels2016In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 16, no 22, p. 2911-2920Article in journal (Refereed)
    Abstract [en]

    The molecular composition of synaptic signal transduction machineries shapes synaptic neurotransmission. The repertoire of receptors, transporters and channels (RTCs) comprises major signaling events in the brain. RTCs are conventionally studied by candidate immunohistochemistry and biochemistry, which are low throughput with resolution greatly affected by available immunoreagents and membrane interference. Therefore, a comprehensive resource of synaptic brain RTCs is still lacking. In particular, studies on the detergent-soluble synaptosomal fraction, known to contain transporters and channels, are limited. We, therefore, performed sub-synaptosomal fractionation of rat cerebral cortex, followed by trypsin/chymotrypsin sequential digestion of a detergent-soluble synaptosomal fraction and a postsynaptic density preparation, stable-isotope tryptic peptide labeling and liquid chromatography mass spectrometry. Based on the current study, a total of 4784 synaptic proteins were submitted to the ProteomExchange database (PXD001948), including 274 receptors, 394 transporters/channels and 1377 transmembrane proteins. Function-based classification assigned 1781 proteins as probable drug targets with 834 directly linked to brain disorders. The analytical approach identified 499 RTCs that are not listed in the largest, curated database for synaptosomal proteins (SynProt). This is a threefold RTC increase over all other data collected to date. Taken together, we present a protein discovery resource that can serve as a benchmark for future molecular interrogation of synaptic connectivity.

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