Proteomic analysis of the presynaptic active zone from APP/APLP mutant mice
Final Report Abstract
Synaptic vesicles docked to the presynaptic plasma membrane comprise the exocytosis-competent subpopulation of vesicles at the active zone. Employing subcellular fractionation of synaptosomes derived from mouse brain and a monoclonal antibody directed against an integral vesicle membrane protein, we could immunoisolate synaptic vesicles together with the attached presynaptic plasma membrane. All three members of the APP family are present in the immunoisolated active zone fraction. This is particularly interesting, as at the neuromuscular junction APP and APLP2 are involved in the control of synaptic vesicle release, with APPsα-DM mice showing reduced quantal content and a pronounced reduction in the readily releasable pool. In order to gain insight into the proteinaceous interactions and possible function of APP proteins it is our goal is to compare the proteome of the presynaptic active zone of APP/APLP-knockout mice to that of controls. In addition, we will investigate if there are differences in the posttranslational modifications of presynaptic proteins following induced synaptic activity between littermate controls and APP/APLP mutant mice. Differences in the protein profile will be analyzed and the functional consequences evaluated by a joint effort within the consortium.
Publications
- (2010) Electrophysiological characterization of ATPases in native synaptic vesicles and synaptic plasma membranes. Biochem J 427:151-159
Obrdlik P, Diekert K, Watzke N, Keipert C, Pehl U, Brosch C, Boehm N, Bick I, Ruitenberg M, Volknandt W, Kelety B
- (2010) Labeling elastase digests with TMT: Informational gain by identification of poorly detectable peptides with MALDI-TOF/TOF mass spectrometry. Proteomics 1:3905–3909
Baeumlisberger D; Arrey TN, Rietschel B, Rohmer M, Papasotiriou DG, Mueller BF, Beckhaus T, Karas M
- (2011) Simple Dual-Spotting Procedure Enhances nLC–MALDI MS/MS Analysis of Digests with Less Specific Enzymes. J. Proteome Res 10:2889–2894
Baeumlisberger D, Rohmer M, Arrey TN, Mueller BF, Beckhaus T, Bahr U, Barka G, Karas, M
- (2011) SV31 is a Zn2+-binding synaptic vesicle protein. J Neurochem 118:558-579
Barth J, Zimmermann H, Volknandt W
- (2012) Enzyme-cleavable tandem peptides for quantitative studies in MS-based proteomics. Proteomics 12:3470-3474
Winter D, Hung CW, Jaskolla TW, Karas M, Lehmann WD
- (2012) Perfusion reversed-phase high-performance liquid chromatography for protein separation from detergent-containing solutions: An alternative to gel-based approaches. Anal Biochem 424:97-107
Gorka J, Rohmer M, Bornemann S, Papasotiriou DG, Baeumlisberger D, Arrey TN, Bahr U, Karas M
