title: Identification of a multifunctional docking site on the catalytic unit of phosphodiesterase-4 (PDE4) that is utilised by multiple interaction partners.
abstract: Cyclic AMP (cAMP)-specific phosphodiesterase-4 (PDE4) enzymes underpin compartmentalised cAMP signalling by localising to distinct signalling complexes. PDE4 long isoforms can be phosphorylated by mitogen-activated protein kinase-activated protein kinase 2 (MK2), which attenuates activation of such enzymes through their phosphorylation by protein kinase A. Here we show that MK2 interacts directly with PDE4 long isoforms and define the sites of interaction. One is a unique site that locates within the regulatory upstream conserved region 1 (UCR1) domain and contains a core Phe141, Leu142 and Tyr143 (FLY) cluster (PDE4A5 numbering). Located with the second site is a critical core Phe693, Glu694, Phe695 (FQF) motif that is also employed in the sequestering of PDE4 long forms by an array of other signalling proteins, including the signalling scaffold β-arrestin, the tyrosyl kinase Lyn, the SUMOylation E2 ligase UBC9, the dynein regulator Lis1 (PAFAH1B1) and the protein kinase Erk. We propose that the FQF motif lies at the heart of a multifunctional docking (MFD) site located within the PDE4 catalytic unit. It is clear from our data that, as well as aiding fidelity of interaction, the MFD site confers exclusivity of binding between PDE4 and a single specific partner protein from the cohort of signalling proteins whose interaction with PDE4 involves the FQF motif.
authors.affiliation: Institute of Cardiovascular and Medical Science, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, U.K.
authors.forename: Kirsty F
authors.initials: KF
authors.lastname: Houslay
authors.affiliation: Institute of Cardiovascular and Medical Science, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, U.K.
authors.forename: Frank
authors.initials: F
authors.lastname: Christian
authors.affiliation: Institute of Cardiovascular and Medical Science, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, U.K.
authors.forename: Ruth
authors.initials: R
authors.lastname: MacLeod
authors.affiliation: Institute of Chemical Sciences, Heriott-Watt University, Edinburgh, U.K.|Mironid Ltd, BioCity Scotland, Bo'Ness Road, Newhouse, North Lanarkshire ML1 5UH, U.K.
authors.forename: David R
authors.initials: DR
authors.lastname: Adams
authors.affiliation: Mironid Ltd, BioCity Scotland, Bo'Ness Road, Newhouse, North Lanarkshire ML1 5UH, U.K.|Institute of Pharmaceutical Sciences, King's College London, London SE1 9NH, U.K.
authors.forename: Miles D
authors.initials: MD
authors.lastname: Houslay
authors.affiliation: Institute of Cardiovascular and Medical Science, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, U.K. george.baillie@glasgow.ac.uk.
authors.forename: George S
authors.initials: GS
authors.lastname: Baillie
mesh_terms: D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs; D000818:Animals; D019556:COS Cells; D020134:Catalytic Domain; D002522:Chlorocebus aethiops; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D062105:Molecular Docking Simulation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000071557:beta-Arrestins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019061:src-Family Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000098843:Ubiquitin-Conjugating Enzyme UBC9; D000099190:MAP-Kinase-Activated Kinase 2
chemical_list: D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; D017868:Cyclic AMP-Dependent Protein Kinases; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D047908:Intracellular Signaling Peptides and Proteins; D008869:Microtubule-Associated Proteins; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D017346:Protein Serine-Threonine Kinases; D011994:Recombinant Proteins; D044763:Ubiquitin-Conjugating Enzymes; D000071557:beta-Arrestins; D019061:src-Family Kinases; D000098843:Ubiquitin-Conjugating Enzyme UBC9; D000099190:MAP-Kinase-Activated Kinase 2; C060902:lyn protein-tyrosine kinase; C535150:MAPK1 protein, human; C084102:PAFAH1B1 protein, human; C517355:PDE4A protein, human
keywords: cAMP; cyclic nucleotide phosphodiesterases; protein–protein interactions