Structure of the catalytic domain of human phosphodiesterase 5 with bound drug molecules

Phosphodiesterases (PDEs) are a superfamily of enzymes that degrade the intracellular second messengers cyclic AMP and cyclic GMP. As essential regulators of cyclic nucleotide signalling with diverse physiological functions, PDEs are drug targets for the treatment of various diseases, including heart failure, depression, asthma, inflammation and erectile dysfunction. Of the 12 PDE gene families, cGMP-specific PDE5 carries out the principal cGMP-hydrolysing activity in human corpus cavernosum tissue. It is well known as the target of sildenafil citrate (Viagra) and other similar drugs for the treatment of erectile dysfunction. Despite the pressing need to develop selective PDE inhibitors as therapeutic drugs, only the cAMP-specific PDE4 structures are currently available. Here we present the three-dimensional structures of the catalytic domain (residues 537–860) of human PDE5 complexed with the three drug molecules sildenafil, tadalafil (Cialis) and vardenafil (Levitra). These structures will provide opportunities to design potent and selective PDE inhibitors with improved pharmacological profiles.

[1]  Melvin I. Simon,et al.  Diversity of G proteins in signal transduction , 1991, Science.

[2]  J. Corbin,et al.  9 – Design and Synthesis of Xanthines and Cyclic GMP Analogues as Potent Inhibitors of PDE5 , 1996 .

[3]  T. Schwartz,et al.  Conversion of antagonist-binding site to metal-ion site in the tachykinin NK-1 receptor , 1995, Nature.

[4]  J. Corbin,et al.  PHARMACOLOGY OF PHOSPHODIESTERASE‐5 INHIBITORS , 2002, International journal of clinical practice.

[5]  Thomas C. Terwilliger,et al.  Automated MAD and MIR structure solution , 1999, Acta crystallographica. Section D, Biological crystallography.

[6]  J. Pouysségur,et al.  Pertussis toxin inhibits thrombin‐induced activation of phosphoinositide hydrolysis and Na+/H+ exchange in hamster fibroblasts. , 1986, The EMBO journal.

[7]  J. Peterson,et al.  Effects of extracellular pH on tumour necrosis factor-alpha production by resident alveolar macrophages. , 2001, Clinical science.

[8]  S. Doublié.  Preparation of selenomethionyl proteins for phase determination. , 1997, Methods in enzymology.

[9]  S. Carrier.  Pharmacology of phosphodiesterase 5 inhibitors. , 2003, The Canadian journal of urology.

[10]  J. Beavo,et al.  Regulation of cAMP and cGMP signaling: new phosphodiesterases and new functions. , 2000, Current opinion in cell biology.

[11]  M. Berridge,et al.  Lithium amplifies agonist-dependent phosphatidylinositol responses in brain and salivary glands. , 1982, The Biochemical journal.

[12]  Roland L. Dunbrack,et al.  Prediction of protein side-chain rotamers from a backbone-dependent rotamer library: a new homology modeling tool. , 1997, Journal of molecular biology.

[13]  A I Basbaum,et al.  Impaired nociception and pain sensation in mice lacking the capsaicin receptor. , 2000, Science.

[14]  David P. Rotella,et al.  INHIBITORS: CURRENT STATUS AND POTENTIAL APPLICATIONS , 2002 .

[15]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[16]  Michael J. Berridge,et al.  Inositol phosphates and cell signalling , 1989, Nature.

[17]  T. Torphy.  Phosphodiesterase isozymes: molecular targets for novel antiasthma agents. , 1998, American journal of respiratory and critical care medicine.

[18]  K. Seuwen,et al.  Stem cell characteristics of human trabecular bone-derived cells. , 2002, Bone.

[19]  U. Lerner,et al.  Characterization of bradykinin receptors in a human osteoblastic cell line , 2002, Regulatory Peptides.

[20]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[21]  K. Palczewski,et al.  Crystal Structure of Rhodopsin: A G‐Protein‐Coupled Receptor , 2002, Chembiochem : a European journal of chemical biology.

[22]  Thomas C. Terwilliger,et al.  Electronic Reprint Biological Crystallography Maximum-likelihood Density Modification , 2022 .

[23]  S. Jin,et al.  Cyclic nucleotide phosphodiesterases and their role in endocrine cell signaling , 2002, Trends in Endocrinology & Metabolism.

[24]  M. Conti,et al.  Recent progress in understanding the hormonal regulation of phosphodiesterases. , 1995, Endocrine reviews.

[25]  J. Beavo,et al.  Cyclic nucleotide phosphodiesterases: functional implications of multiple isoforms. , 1995, Physiological reviews.

[26]  T. Arnett,et al.  pH dependence of bone resorption: mouse calvarial osteoclasts are activated by acidosis. , 2001, American journal of physiology. Endocrinology and metabolism.

[27]  P. Anand,et al.  TRPV3 is a temperature-sensitive vanilloid receptor-like protein , 2002, Nature.

[28]  Kornelia Polyak,et al.  Mechanism of CDK activation revealed by the structure of a cyclinA-CDK2 complex , 1995, Nature.

[29]  D. Bushinsky,et al.  Prostaglandins regulate acid-induced cell-mediated bone resorption. , 2000, American journal of physiology. Renal physiology.

[30]  O. Lichtarge,et al.  Rhodopsin activation blocked by metal-ion-binding sites linking transmembrane helices C and F , 1996, Nature.

[31]  D. Bushinsky.  Acid-base imbalance and the skeleton , 2001, European journal of nutrition.

[32]  J. Beavo,et al.  PDE5 is converted to an activated state upon cGMP binding to the GAF A domain , 2003, The EMBO journal.

[33]  H. Ishizaka,et al.  Coronary arteriolar dilation to acidosis: role of ATP-sensitive potassium channels and pertussis toxin-sensitive G proteins. , 1999, Circulation.

[34]  S. Burley,et al.  Crystal structure of a TFIIB–TBP–TATA-element ternary complex , 1995, Nature.

[35]  C. Leslie,et al.  Regulation of arachidonic acid release and cytosolic Phospholipase A2 activation , 1999, Journal of leukocyte biology.

[36]  Y. Zhao,et al.  Atomic structure of PDE4: insights into phosphodiesterase mechanism and specificity. , 2000, Science.

[37]  Y. Salomon.  Adenylate cyclase assay. , 1979, Advances in cyclic nucleotide research.

[38]  Joseph Markowitz,et al.  Crystal structure of phosphodiesterase 4D and inhibitor complex1 , 2002, FEBS letters.

[39]  P. Reeh,et al.  Tissue acidosis in nociception and pain. , 1996, Progress in brain research.

[40]  J. Corbin,et al.  Cyclic GMP Phosphodiesterase-5: Target of Sildenafil* , 1999, The Journal of Biological Chemistry.

[41]  Z. Otwinowski,et al.  Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[42]  J. Corbin,et al.  Potential Roles of Conserved Amino Acids in the Catalytic Domain of the cGMP-binding cGMP-specific Phosphodiesterase (PDE5)* , 1998, The Journal of Biological Chemistry.

[43]  E. Goetzl,et al.  Cloning, sequencing and tissue distribution of two related G protein‐coupled receptor candidates expressed prominently in human lung tissue , 1995, FEBS letters.

[44]  P. Kollman,et al.  A Second Generation Force Field for the Simulation of Proteins, Nucleic Acids, and Organic Molecules , 1995 .