title: Proteomic analysis of mouse brain microsomes: identification and bioinformatic characterization of endoplasmic reticulum proteins in the mammalian central nervous system.
abstract: The endoplasmic reticulum (ER) is the main source for the storage and release of intracellular calcium in neurons and, thus, contributes to the functionality of a diverse set of pathways that control critical aspects of central nervous system function including but not limited to gene expression, neurotransmission, learning, and memory. ER-derived proteins obtained after subcellular fractionation of mouse brain homogenate were digested with trypsin and the corresponding peptides fractionated by strong cation exchange chromatography followed by LC-MS/MS analysis on a hybrid linear ion trap--Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. A comprehensive catalogue representing 1914 proteins was generated from this particular proteomic analysis using identification criteria that corresponded to a false positive identification rate of 0.4%. Various molecular functions and biological processes relevant to the ER were identified upon gene ontology (GO)-based analysis including pathways associated with molecular transport, protein trafficking and localization, and cell signaling. Comparison of the 2D-LC-MS/MS results with those obtained from shotgun LC-MS/MS analyses demonstrated that most molecular functions and biological processes were represented via GO analysis using either methodology. Results from this comparison as well as a focused investigation into components of calcium-mediated signaling in the mouse brain ER are also presented.
authors.affiliation: Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Forth Worth, Texas 76107, USA.
authors.forename: Stanley M
authors.initials: SM
authors.lastname: Stevens
authors.forename: R Scott
authors.initials: RS
authors.lastname: Duncan
authors.forename: Peter
authors.initials: P
authors.lastname: Koulen
authors.forename: Laszlo
authors.initials: L
authors.lastname: Prokai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002490:Central Nervous System / Q000378:metabolism*; D019295:Computational Biology; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D016247:Information Storage and Retrieval; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008861:Microsomes / Q000378:metabolism*; D040901:Proteomics; D017550:Spectroscopy, Fourier Transform Infrared; D053719:Tandem Mass Spectrometry