Intracellular Compartmentalization of PDE4 Cyclic AMP-Specific Phosphodiesterases

• Published in: Methods (San Diego, Calif.) Vol. 14; no. 1; pp. 65 - 79
• Main Authors: 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.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.1998
• Subjects: 3',5'-Cyclic-AMP Phosphodiesterases - analysis; Alternative Splicing - genetics; Animals; COS Cells; Cyclic Nucleotide Phosphodiesterases, Type 4; DNA Primers - chemistry; Fluorescent Antibody Technique; Gene Expression - genetics; Isoenzymes - analysis; Membrane Proteins - analysis; Microscopy, Confocal; Mutagenesis, Site-Directed - genetics; Plasmids - genetics; Protein Biosynthesis - genetics; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Sequence Deletion; src Homology Domains - genetics; Transcription, Genetic - genetics; Transfection - genetics
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1006/meth.1997.0566

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 havingsrchomology 3 (SH3) domains, and generation of chimeric constructs of these N-terminal regions with proteins that are normally expressed in the cytosol.