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Purification of bovine neurosecretory granule membranes by density gradient centrifugation
Authors:C B González  C E Caorsi  W G North  O T Berrios
Institution:1. Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA), CONICET, Facultad de Ciencias, Exactas, UNLP, CIC, Calle 47 y 116, B1900AJJ La Plata, Argentina;2. Centro de Investigación y Desarrollo en Ciencias Aplicadas (CINDECA-CONICET-CIC-UNLP), Calle 47 N° 257, B1900AJJ La Plata, Argentina;3. Facultad de Ingeniería, UNLP, Calle 47 y 1, B1900AJJ La Plata, Argentina;1. Center for Brain and Cognition, Computational Neuroscience Group, Department of Information and Communication Technologies, Universitat Pompeu Fabra, Spain;2. Clinical Neurophysiology Group, University of Twente, Enschede, The Netherlands;3. Department of Neurology, Amsterdam UMC, Amsterdam, the Netherlands;4. Sir Peter Mansfield Imaging Centre, School of Physics, University of Nottingham, Nottingham, United Kingdom;5. Department of Mathematics, Faculty of Science, Vrije Universiteit Amsterdam, Amsterdam, the Netherlands;6. Sir Peter Mansfield Imaging Centre, School of Medicine, University of Nottingham, United Kingdom;7. NIHR Biomedical Research Centre, University of Nottingham, Nottingham University Hospitals NHS Trust, Nottingham, UK;8. Centre for Eudaimonia and Human Flourishing, Linacre College, University of Oxford, Oxford, UK;9. Center for Music in the Brain, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark;10. Department of Psychiatry, University of Oxford, Oxford, UK;11. Institució Catalana de la Recerca i Estudis Avançats (ICREA), Barcelona, Spain;12. Department of Neuropsychology, Max Planck Institute for Human Cognitive and Brain Sciences, Leipzig, Germany
Abstract:A procedure for the subfractionation of neurosecretory granules into membrane and content components is described. The procedure involves the hypotonic lysis of the secretory granule fraction and further purification of the membranes by centrifugation through a discontinuous sucrose gradient. The neurosecretory granule membranes represented 5.2% of the total proteins of the neurosecretory granule fraction and were highly enriched in cytochrome b561. Electron microscopic analysis of the purified membranes showed vesicles devoid of electrodense content.
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