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Mass spectrometric analysis of synaptosomal membrane preparations for the determination of brain receptors, transporters and channels
Author(s) -
Sialana Fernando J.,
Gulyassy Peter,
Májek Peter,
Sjöstedt Evelina,
Kis Viktor,
Müller André C.,
Rudashevskaya Elena L.,
Mulder Jan,
Bennett Keiryn L.,
Lubec Gert
Publication year - 2016
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201600234
Subject(s) - neurotransmission , transporter , transmembrane protein , receptor , chemistry , biochemistry , biology , gene
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.