title: Proteome Alterations of Hippocampal Cells Caused by Clostridium botulinum C3 Exoenzyme.
abstract: C3bot from Clostridium botulinum is a bacterial mono-ADP-ribosylating enzyme, which transfers an ADP-ribose moiety onto the small GTPases Rho A/B/C. C3bot and the catalytic inactive mutant (C3E174Q) cause axonal and dendritic growth as well as branching in primary hippocampal neurons. In cultured murine hippocampal HT22 cells, protein abundances were analyzed in response to C3bot or C3E174Q treatment using a shotgun proteomics approach. Proteome analyses were performed at four time points over 6 days. More than 4000 protein groups were identified at each time point and quantified in triplicate analyses. On day one, 46 proteins showed an altered abundance, and after 6 days, more than 700 proteins responded to C3bot with an up- or down-regulation. In contrast, C3E174Q had no provable impact on protein abundance. Protein quantification was verified for several proteins by multiple reaction monitoring. Data analysis of altered proteins revealed different cellular processes that were affected by C3bot. They are particularly involved in mitochondrial and lysosomal processes, adhesion, carbohydrate and glucose metabolism, signal transduction, and nuclear proteins of translation and ribosome biogenesis. The results of this study gain novel insights into the function of C3bot in hippocampal cells.
authors.affiliation: Institute of Toxicology, Hannover Medical School , Carl-Neuberg-Str.1, 30625 Hannover, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Schröder
authors.affiliation: Institute of Toxicology, Hannover Medical School , Carl-Neuberg-Str.1, 30625 Hannover, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Rohrbeck
authors.affiliation: Institute of Toxicology, Hannover Medical School , Carl-Neuberg-Str.1, 30625 Hannover, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Just
authors.affiliation: Institute of Toxicology, Hannover Medical School , Carl-Neuberg-Str.1, 30625 Hannover, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Pich
mesh_terms: D036002:ADP Ribose Transferases / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D000818:Animals; D001905:Botulinum Toxins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D050260:Carbohydrate Metabolism / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D003014:Clostridium botulinum / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks / Q000187:drug effects*; D005947:Glucose / Q000378:metabolism; D006624:Hippocampus / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008247:Lysosomes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D058977:Molecular Sequence Annotation; D009154:Mutation; D009474:Neurons / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001678:Organelle Biogenesis; D061251:Primary Cell Culture; D014176:Protein Biosynthesis / Q000187:drug effects; D020543:Proteome / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D012270:Ribosomes / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D009687:Nuclear Proteins; D020543:Proteome; D011994:Recombinant Proteins; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D001905:Botulinum Toxins; D005947:Glucose
keywords: C3 exoenzyme; Clostridium botulinum; SILAC; shotgun proteomics