Intracellular Localization of 2',3'-Cyclic Nucleotide 3'-Phosphodiesterase in a Neuronal Cell Line as Examined by Immunofluorescence and Cell Fractionation |
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Authors: | Hans Werner Mülller Patric A Clapshaw Wilfried Seifert |
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Institution: | Friedrich-Miescher-Laboratorium der Max-Planck-Gesellschaft, Tübingen, F.R.G. |
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Abstract: | The enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase, EC 3.1.4.37) occurs not only in myelin fractions and glial cells, but can also be shown to be present in a CNS cell line of neuronal origin (B104). Direct immunofluorescence microscopy of B104 cells with fluorescein isothiocyanate-conjugated rabbit anti-CNPase antibodies shows a discrete and specific intracytoplasmic location of CNPase. Fractionation of the cells was performed by differential centrifugation of a cell homogenate and continuous sucrose density-gradient centrifugation. As monitored by marker enzyme activities, CNPase seems to be associated with endoplasmic reticulum membranes. |
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Keywords: | CNPase Neuronal cell line Myelin Immunofluorescence Cell fractionation Endoplasmic reticulum |
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