PMID- 25865307 OWN - NLM STAT- MEDLINE DCOM- 20160412 LR - 20220309 IS - 1876-7737 (Electronic) IS - 1874-3919 (Linking) VI - 123 DP - 2015 Jun 18 TI - Proteomic analysis of the palmitoyl protein thioesterase 1 interactome in SH-SY5Y human neuroblastoma cells. PG - 42-53 LID - S1874-3919(15)00160-8 [pii] LID - 10.1016/j.jprot.2015.03.038 [doi] AB - Neuronal ceroid lipofuscinoses (NCL) are a group of inherited progressive childhood disorders, characterized by early accumulation of autofluorescent storage material in lysosomes of neurons or other cells. Clinical symptoms of NCL include: progressive loss of vision, mental and motor deterioration, epileptic seizures and premature death. CLN1 disease (MIM#256730) is caused by mutations in the CLN1 gene, which encodes palmitoyl protein thioesterase 1 (PPT1). In this study, we utilised single step affinity purification coupled to mass spectrometry (AP-MS) to unravel the in vivo substrates of human PPT1 in the brain neuronal cells. Protein complexes were isolated from human PPT1 expressing SH-SY5Y stable cells, subjected to filter-aided sample preparation (FASP) and analysed on a Q Exactive Hybrid Quadrupole-Orbitrap mass spectrometer. A total of 23 PPT1 interacting partners (IP) were identified from label free quantitation of the MS data by SAINT platform. Three of the identified PPT1 IP, namely CRMP1, DBH, and MAP1B are predicted to be palmitoylated. Our proteomic analysis confirmed previously suggested roles of PPT1 in axon guidance and lipid metabolism, yet implicates the enzyme in novel roles including: involvement in neuronal migration and dopamine receptor mediated signalling pathway. BIOLOGICAL SIGNIFICANCE: The significance of this work lies in the unravelling of putative in vivo substrates of human CLN1 or PPT1 in brain neuronal cells. Moreover, the PPT1 IP implicate the enzyme in novel roles including: involvement in neuronal migration and dopamine receptor mediated signalling pathway. CI - Copyright (c) 2015 Elsevier B.V. All rights reserved. FAU - Scifo, Enzo AU - Scifo E AD - Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, University of Helsinki, Helsinki, Finland; Doctoral Program Brain & Mind, University of Helsinki, Helsinki, Finland. Electronic address: enzo.scifo@helsinki.fi. FAU - Szwajda, Agnieszka AU - Szwajda A AD - Institute for Molecular Medicine (FIMM), University of Helsinki, Helsinki, Finland. FAU - Soliymani, Rabah AU - Soliymani R AD - Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, University of Helsinki, Helsinki, Finland. FAU - Pezzini, Francesco AU - Pezzini F AD - Department of Neurological and Movement Sciences, University of Verona, Verona, Italy. FAU - Bianchi, Marzia AU - Bianchi M AD - Department of Neurological and Movement Sciences, University of Verona, Verona, Italy; Unit for Neuromuscular and Neurodegenerative Disorders, Laboratory of Molecular Medicine, Bambino Gesu Children's Hospital, IRCCS, Rome, Italy. FAU - Dapkunas, Arvydas AU - Dapkunas A AD - Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, University of Helsinki, Helsinki, Finland. FAU - Debski, Janusz AU - Debski J AD - Mass Spectrometry Laboratory, Department of Biophysics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland. FAU - Uusi-Rauva, Kristiina AU - Uusi-Rauva K AD - Folkhalsan Institute of Genetics, Helsinki, Finland; National Institute for Health and Welfare, Public Health Genomics Unit, Helsinki, Finland. FAU - Dadlez, Michal AU - Dadlez M AD - Mass Spectrometry Laboratory, Department of Biophysics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland. FAU - Gingras, Anne-Claude AU - Gingras AC AD - Centre for Systems Biology, Samuel Lunenfeld Research Institute at Mount Sinai Hospital, Toronto, Canada; Department of Molecular Genetics, University of Toronto, Ontario, Canada. FAU - Tyynela, Jaana AU - Tyynela J AD - Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, University of Helsinki, Helsinki, Finland. FAU - Simonati, Alessandro AU - Simonati A AD - Department of Neurological and Movement Sciences, University of Verona, Verona, Italy. FAU - Jalanko, Anu AU - Jalanko A AD - Institute for Molecular Medicine (FIMM), University of Helsinki, Helsinki, Finland; National Institute for Health and Welfare, Public Health Genomics Unit, Helsinki, Finland. FAU - Baumann, Marc H AU - Baumann MH AD - Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, University of Helsinki, Helsinki, Finland. FAU - Lalowski, Maciej AU - Lalowski M AD - Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, University of Helsinki, Helsinki, Finland; Folkhalsan Institute of Genetics, Helsinki, Finland. Electronic address: maciej.lalowski@helsinki.fi. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150410 PL - Netherlands TA - J Proteomics JT - Journal of proteomics JID - 101475056 RN - 0 (Membrane Proteins) RN - EC 3.1.2.- (Thiolester Hydrolases) RN - EC 3.1.2.22 (PPT1 protein, human) SB - IM MH - Axons/metabolism MH - Brain/metabolism MH - Cell Line, Tumor MH - Cell Movement MH - Energy Metabolism MH - *Gene Expression Regulation, Neoplastic MH - Glycosylation MH - HEK293 Cells MH - Humans MH - Lysosomes/metabolism MH - Mass Spectrometry MH - Membrane Proteins/genetics/*metabolism MH - Microscopy, Fluorescence MH - Mitochondria/physiology MH - Mutation MH - Neuroblastoma/*metabolism MH - Neuronal Ceroid-Lipofuscinoses/metabolism MH - Neurons/metabolism MH - Open Reading Frames MH - Proteomics/*methods MH - Signal Transduction MH - Thiolester Hydrolases OTO - NOTNLM OT - Affinity purification OT - CLN1 disease OT - Interactome OT - Mass spectrometry OT - Neuronal ceroid lipofuscinoses OT - Palmitoyl protein thioesterase 1 EDAT- 2015/04/14 06:00 MHDA- 2016/04/14 06:00 CRDT- 2015/04/14 06:00 PHST- 2014/11/28 00:00 [received] PHST- 2015/03/12 00:00 [revised] PHST- 2015/03/31 00:00 [accepted] PHST- 2015/04/14 06:00 [entrez] PHST- 2015/04/14 06:00 [pubmed] PHST- 2016/04/14 06:00 [medline] AID - S1874-3919(15)00160-8 [pii] AID - 10.1016/j.jprot.2015.03.038 [doi] PST - ppublish SO - J Proteomics. 2015 Jun 18;123:42-53. doi: 10.1016/j.jprot.2015.03.038. Epub 2015 Apr 10.