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Intracellular Localization of 2′,3′‐Cyclic Nucleotide 3′‐Phosphodiesterase in a Neuronal Cell Line as Examined by Immunofluorescence and Cell Fractionation
Author(s) -
Mülller Hans Werner,
Clapshaw Patric A.,
Seifert Wilfried
Publication year - 1981
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1981.tb04482.x
Subject(s) - intracellular , immunofluorescence , cell fractionation , cell culture , nucleotide , fractionation , phosphodiesterase , cell , microbiology and biotechnology , cyclic nucleotide , cyclic nucleotide phosphodiesterase , chemistry , biochemistry , biology , enzyme , genetics , antibody , chromatography , gene
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 intra‐cytoplasmic 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.