Intracellular compartmentalization of PDE4 cyclic AMP-specific phosphodiesterases

G. Scotland, M. Beard, S. Erdogan, E. Huston, F. McCallum, S. J. MacKenzie, A. H. Peden, L. Pooley, N. G. Rena, A H Ross, S. J. Yarwood, M. D. Houslay

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    9 Citations (Scopus)

    Abstract

    The PDE4 cyclic AMP-specific phosphodiesterase family comprises a large number of different isoforms encoded by four distinct genes, with additional complexity arising through alternate mRNA splicing. This generates a number of distinct PDE4 isoforms with unique N-terminal regions. The range of such splice variants emanating from the four PDE4 genes appears to be highly conserved across species. One key role for such regions appears to be their potential to target isoforms to specific intracellular sites. Evidence for such a targeting role for these N-terminal regions can be gleaned by a variety of techniques. These include subcellular fractionation, confocal microscopy, binding assays to show association with proteins having src homology 3 (SH3) domains, and generation of chimeric constructs of these N-terminal regions with proteins that are normally expressed in the cytosol.

    Original languageEnglish
    Pages (from-to)65-79
    Number of pages15
    JournalMethods
    Volume14
    Issue number1
    DOIs
    Publication statusPublished - Jan 1998

    Keywords

    • 3',5'-Cyclic-AMP Phosphodiesterases
    • Alternative Splicing
    • Animals
    • COS Cells
    • Cyclic Nucleotide Phosphodiesterases, Type 4
    • DNA Primers
    • Fluorescent Antibody Technique
    • Gene Expression
    • Isoenzymes
    • Membrane Proteins
    • Microscopy, Confocal
    • Mutagenesis, Site-Directed
    • Plasmids
    • Protein Biosynthesis
    • Recombinant Fusion Proteins
    • Sequence Deletion
    • Transcription, Genetic
    • Transfection
    • src Homology Domains

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  • Cite this

    Scotland, G., Beard, M., Erdogan, S., Huston, E., McCallum, F., MacKenzie, S. J., Peden, A. H., Pooley, L., Rena, N. G., Ross, A. H., Yarwood, S. J., & Houslay, M. D. (1998). Intracellular compartmentalization of PDE4 cyclic AMP-specific phosphodiesterases. Methods, 14(1), 65-79. https://doi.org/10.1006/meth.1997.0566