DOCSLIB.ORG

Soluble Guanylate Cyclase B1-Subunit Expression Is Increased in Mononuclear Cells from Patients with Erectile Dysfunction

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Soluble Guanylate Cyclase B1-Subunit Expression Is Increased in Mononuclear Cells from Patients with Erectile Dysfunction International Journal of Impotence Research (2006) 18, 432–437 & 2006 Nature Publishing Group All rights reserved 0955-9930/06 $30.00 www.nature.com/ijir ORIGINAL ARTICLE Soluble guanylate cyclase b1-subunit expression is increased in mononuclear cells from patients with erectile dysfunction PJ Mateos-Ca´ceres1, J Garcia-Cardoso2, L Lapuente1, JJ Zamorano-Leo´n1, D Sacrista´n1, TP de Prada1, J Calahorra2, C Macaya1, R Vela-Navarrete2 and AJ Lo´pez-Farre´1 1Cardiovascular Research Unit, Cardiovascular Institute, Hospital Clı´nico San Carlos, Madrid, Spain and 2Urology Department, Fundacio´n Jime´nez Diaz, Madrid, Spain The aim was to determine in circulating mononuclear cells from patients with erectile dysfunction (ED), the level of expression of endothelial nitric oxide synthase (eNOS), soluble guanylate cyclase (sGC) b1-subunit and phosphodiesterase type-V (PDE-V). Peripheral mononuclear cells from nine patients with ED of vascular origin and nine patients with ED of neurological origin were obtained. Fourteen age-matched volunteers with normal erectile function were used as control. Reduction in eNOS protein was observed in the mononuclear cells from patients with ED of vascular origin but not in those from neurological origin. Although sGC b1-subunit expression was increased in mononuclear cells from patients with ED, the sGC activity was reduced. However, only the patients with ED of vascular origin showed an increased expression of PDE-V. This work shows for the first time that, independently of the aetiology of ED, the expression of sGC b1-subunit was increased in circulating mononuclear cells; however, the expression of both eNOS and PDE-V was only modified in the circulating mononuclear cells from patients with ED of vascular origin. International Journal of Impotence Research (2006) 18, 432–437. doi:10.1038/sj.ijir.3901461; published online 9 March 2006 Keywords: molecular biologic study; risk factor Introduction stimulates soluble guanylate cyclase (sGC) and the activation of sGC elevates intracellular levels of Penile erection is regulated by the relaxation of cGMP, resulting in relaxation of the corporal smooth arteriolar and trabecular smooth muscle cells which muscle, dilatation of the penile arteries, arterioles it is mediated by the nitric oxide (NO)–cyclic and sinusoids, and finally, increased arterial inflow guanosine monophosphate (cGMP) pathway. Nitric and passive compression of the penile venous oxide is a nonadrenergic–noncholinergic neuro- outflow.4,5 Soluble guanylate cyclase is a hetero- transmitter that is released from postgangliomic dimer composed of a large (a1) and a small (b1) parasympathetic (cholinergic) nerves in the penis subunits and both subunits are required to produce and NO is also released from the endothelium.1–3 cGMP.6 Cyclic guanosine monophosphate is catabo- Both neuronal NOS (nNOS) isoform and the lized by phosphodiesterase type-V (PDE-V) that it endothelial NOS (eNOS) participate in cavernosal has been identified in human cavernous tissue and relaxation. plays a relevant role in penile erection. However, it Independently of the NO generating systems, a is now known that other tissues expressed PDE-V.7 reduction in the NO-induced vasorelaxation could Leukocytes also produce NO by an eNOS-like also results from impaired downstream signaling NOS and contain sGC.8,9 However, despite of in the vascular smooth muscle cells. Nitric oxide leukocytes are more accessible cells than the vascular cells contained in the corpus cavernosum, the ability of the leukocytes to express eNOS, sGC and PDE-V in patients with erectile dysfunction (ED) Correspondence: Dr AJ Lo´pez-Farre´, Cardiovascular has not been explored. Therefore, the aim of the Research Unit, 7a Norte, Hospital Clı´nico San Carlos, Profesor Martı´n Lagos s/n, Madrid 28040, Spain. present study was to test if mononuclear cells from E-mail: [email protected] patients with ED of vascular and neurological origin Received 26 July 2005; revised 5 January 2006; accepted have modified the level of expression of either 23 January 2006; published online 9 March 2006 eNOS, sGC and PDE-V. Mononuclear cells and erectile dysfunction PJ Mateos-Ca´ceres et al 433 Materials and methods polyacrylamide gels. Equal amounts of proteins (10 mg per lane) estimated by bicinchonic acid Study protocol reagent (Pierce Rockford, IL, USA) were loaded. Male ED patients ranging in age from 33 to 81 years Western blot analysis was performed with a mono- (mean 55.4) were enrolled in the study. All patients clonal antibody against eNOS (1:1500 Transduction were initially evaluated by taking a detailed sexual Laboratories, Lexington, UK), sGC b1-subunit and medical history and diagnosed of ED as (1:1500 Alexis Laboratories, San Diego, CA, USA) evaluated by the IIEF standardized questionnaire.10 or PDE-V (1:2500 Calbiochem, La Jolla, CA, USA). Six patients did not achieve a full rigid erection, Specific eNOS, sGC b-subunit and PDE-V proteins three patients showed a short-length full erection were detected by enhanced chemoluminiscense and nine patients showed lack of erection. Patients (ECL, Amersham Corp) and evaluated by densito- were divided into two categories according to the metry (Molecular Dynamics). Prestained protein echo-Doppler analysis after intravenous prostaglan- markers (Sigma) were used for molecular mass din E1 infusion. The vascular ED group included determinations. patients (n ¼ 9; mean age 55.6 years) that showed alterations to prostaglandin E1, a vasodilator, during the echo-Doppler examination based on the peak Determination of soluble guanylate cyclase activity systolic velocity and resistance index. In this group, 5 Â 106 mononuclear cells per tube were incubated the patients also showed previous history of vascu- at 371C in RPMI medium containing 0.025% bovine lar risk factors. In this regard, one patient had serum albumin for 15 min. To investigate the history of hypertension, two had hyperlipemia, two functional activity of sGC, mononuclear cells were had ischemic heart disease associated with hyper- incubated in the presence and in the absence of a lipemia and four patients had not vascular risk NO-donor, sodium nitroprusside (10À5 mol/l; SNP) factors. and a PDE inhibitor, 3-isobutil-1-methylxanthine The second group was the neurological ED group (10À4 mol/l; IBMX) that prevent cGMP breakdown. that included patients (n ¼ 9; mean age 56.6 years) Afterwards, mononuclear cells were lysed in 1 mol/l with ED of neurological origin each of whom ClH in ethanol. After having been mixed, the achieved a full rigid erection after prostaglandin E1 homogenates were centrifuged (12 000 g, 15 min at injection. This group included two patients with 41C) and the supernatants were recovered and dried radical prostatectomy, one patient with cortical by speed-vac. Cyclic guanosine monophosphate was atrophy, two patients with diabetes and four measured in acetylated samples with a kit from patients with peripheral neuropathy. Patients taking Amersham International as reported.14 PDE-V inhibitors in the 6 months previous to the study were excluded. Fourteen age-matched (mean age 54.7 years) with normal erectile function were Statistical analysis used as reference for control values. These age- Results are expressed as mean7s.e.m. Comparisons matched patients were going to be operated of were performed by ANOVA. Bonferroni’s correction Cryptorchidism, Varicocele and Hydrocele. All sub- for multiple comparisons was used to determine the jects gave fully informed consent and the study was level of significance of the P-value. Po0.05 was approved by the local Ethic Committee. The study considered significant. was blind for the investigators that determined the molecular parameters and analyzed the results. Blood samples were drawn from a peripheral vein. Mononuclear cells were isolated by Results Ficoll-Hypaque centrifugation and the layer con- taining leukocytes and lymphocytes was removed Endothelial nitric oxide synthase expression and immediately frozen at À801C for molecular in mononuclear cells determinations. A significant reduction in the level of eNOS protein expressed in mononuclear cells obtained from patients with ED was observed compared with those from age-matched controls (Figure 1). Determination of endothelial nitric oxide synthase, When ED patients were divided as ED of vascular soluble guanylate cyclase b1 subunit and and neurological origin, only mononuclear cells phosphodiesterase type-V from the vascular ED patients showed a significant The expression of eNOS, sGC b1-subunit and reduction of the eNOS protein level compared PDE-V proteins was analyzed by Western blot with those from age-matched controls (Figure 2). as described.11,12 In brief, mononuclear cells were Endothelial nitric oxide synthase expression resuspended and solubilized in Laemmli buffer also tended to be reduced in the mononuclear cells containing 2-mercaptoethanol.13 The proteins from patients with ED of neurological origin obtained were separated in denaturing SDS/10% although it did not reach statistical significance International Journal of Impotence Research Mononuclear cells and erectile dysfunction PJ Mateos-Ca´ceres et al 434 100 300 Control Control E.D. E.D. 250 80 * * 200 60 150 40 100 sGC arbitrary units (a.u.) eNOS arbitrary units (a.u.) 20 50 0 0 Figure 1 Bar graph showing the densitometric analysis of the Figure 3 Bar graph showing the densitometric analysis of the Western blots that detected the expression of the endothelial Western blots
Load more

Recommended publications
  • Response to Inhaled Nitric Oxide, but Neither Sodium Nitroprusside Nor Sildenafil, Predicts Survival in Patients With
    Jachec et al., J Clin Exp Cardiolog 2015, 6:6 Clinical & Experimental Cardiology http://dx.doi.org/10.4172/2155-9880.1000376 Research Article Open Access Response to Inhaled Nitric Oxide, But neither Sodium Nitroprusside nor Sildenafil, Predicts Survival in Patients with Dilated Cardiomyopathy Complicated with Pulmonary Hypertension Wojciech Jacheć1*, Celina Wojciechowska2, Andrzej Tomasik2, Damian Kawecki2, Ewa Nowalany-Kozielska2 and Jan Wodniecki2 1II Katedra i Oddział Kliniczny Kardiologii w Zabrzu Śląskiego Uniwersytetu Medycznego w Katowicach, ul. Skłodowskiej 10, 41-800 Zabrze, Polska 2Department of Cardiology in Zabrze, Medical University of Silesia in Katowice, Poland *Corresponding author: Wojciech Jacheć, II Katedra i Oddział Kliniczny Kardiologii w Zabrzu Śląskiego Uniwersytetu Medycznego w Katowicach, ul. Skłodowskiej 10, 41-800 Zabrze, Polska, Tel: +48 32 373 23 72; Fax: +48 32 271 10 10; E-mail: [email protected] Received date: May 26, 2015, Accepted date: Jun 25, 2015, Published date: Jun 29, 2015 Copyright: ©2015 Jacheć W. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Introduction: Pulmonary hypertension in patients with dilated cardiomyopathy is associated with higher mortality. Objectives: The aim of the study was to assess the predictive value of the vasodilator response to three different drugs, sodium nitroprusside,
    [Show full text]
  • Identi®Cation and Role of Adenylyl Cyclase in Auxin Signalling in Higher
    letters to nature + + + P.P. thank the Academy of Finland and the Deutsche Forschungsgemeinschaft, respectively, for ®nancial CO , 53), 77 (C6H5 , 60), 73 (TMSi , 84); 6-methyl-4-hydroxy-2-pyrone: RRt 0.35, 198 (M+, 18), 183 ([M-Me]+, 16), 170 ([M-CO]+, 54), 155 ([M-CO-Me]+, support. + + Correspondence and requests for materials should be addressed to J.S. (e-mail: [email protected] 15), 139 ([M-Me-CO2] , 10), 127 ([M-Me-2CO] , 13), 99 (12), 84 (13), 73 + + freiburg.de). (TMSi , 100), 43 (CH3CO , 55). The numbers show m/z values, and the key fragments and their relative intensities are indicated in parentheses. Received 4 August; accepted 14 October 1998. erratum 1. Helariutta, Y. et al. Chalcone synthase-like genes active during corolla development are differentially expressed and encode enzymes with different catalytic properties in Gerbera hybrida (Asteraceae). Plant Mol. Biol. 28, 47±60 (1995). 2. Helariutta, Y. et al. Duplication and functional divergence in the chalcone synthase gene family of 8 Asteraceae: evolution with substrate change and catalytic simpli®cation. Proc. Natl Acad. Sci. USA 93, Crystal structure of the complex 9033±9038 (1996). 3. Thaisrivongs, S. et al. Structure-based design of HIV protease inhibitors: 5,6-dihydro-4-hydroxy-2- of the cyclin D-dependent pyrones as effective, nonpeptidic inhibitors. J. Med. Chem. 39, 4630±4642 (1996). 4. Hagen, S. E. et al. Synthesis of 5,6-dihydro-4-hydroxy-2-pyrones as HIV-1 protease inhibitors: the kinase Cdk6 bound to the profound effect of polarity on antiviral activity. J. Med. Chem.
    [Show full text]
  • Acute Effect of Sodium Nitroprusside on Microvascular Dysfunction In
    Clinical Studies Acute Effect of Sodium Nitroprusside on Microvascular Dysfunction in Patients Who Underwent Percutaneous Coronary Intervention for Acute ST-segment Elevation Myocardial Infarction Kotaro Morimoto,1, 2 MD, Shigenori Ito,2 MD, Kosuke Nakasuka,2 MD, Satoru Sekimoto,2 MD, Kazuyuki Miyata,2 MD, Masahiko Inomata,2 MD, Takayuki Yoshida,2 MD, Nozomu Tamai,2 MD, Tomoaki Saeki,2 MD, Shin Suzuki,2 MD, Yoshimasa Murakami,2 MD, Koichi Sato,2 MD, Akihiro Morino,3 CE, and Yoshiyuki Shimizu,3 CE Summary Even in the era of thrombus aspiration and distal protection for ST-segment elevation acute myocardial infarction (STEMI), microvascular dysfunction does exist and improvement of microvascular dysfunction can improve the progno- sis and/or left ventricular dysfunction. We evaluated the acute effects of nitroprusside (NTP) on coronary microvascular injury that occurred after primary percutaneous coronary intervention (PCI) for STEMI in 18 patients. The final Throm- bolysis in Myocardial Infarction trial (TIMI) flow grade after PCI was 3 in 17 patients and 2 in 1 patient. The index of microcirculatory resistance (IMR) was improved significantly from 76 ± 42 to 45 ± 37 (P = 0.0006) by intracoronary NTP administration. IMR improved to the normal range (IMR < 30) in 9 patients (50%). Higher TIMI flow grade and lower IMR at baseline were observed more frequently in patients whose IMR recovered to normal range after NTP ad- ministration. NTP improved the microcirculatory dysfunction at the acute phase in patients who underwent PCI for STEMI and
    [Show full text]
  • Use of Computational Biochemistry for Elucidating Molecular Mechanisms of Nitric Oxide Synthase
    Use of Computational Biochemistry for Elucidating Molecular Mechanisms of Nitric Oxide Synthase Bignon, Emmanuelle; Rizza, Salvatore; Filomeni, Giuseppe; Papaleo, Elena Published in: Computational and Structural Biotechnology Journal DOI: 10.1016/j.csbj.2019.03.011 Publication date: 2019 Document version Publisher's PDF, also known as Version of record Citation for published version (APA): Bignon, E., Rizza, S., Filomeni, G., & Papaleo, E. (2019). Use of Computational Biochemistry for Elucidating Molecular Mechanisms of Nitric Oxide Synthase. Computational and Structural Biotechnology Journal, 17, 415- 429. https://doi.org/10.1016/j.csbj.2019.03.011 Download date: 02. Oct. 2021 Computational and Structural Biotechnology Journal 17 (2019) 415–429 Contents lists available at ScienceDirect journal homepage: www.elsevier.com/locate/csbj Mini Review Use of Computational Biochemistry for Elucidating Molecular Mechanisms of Nitric Oxide Synthase Emmanuelle Bignon a,⁎, Salvatore Rizza b, Giuseppe Filomeni b,c, Elena Papaleo a,d,⁎⁎ a Computational Biology Laboratory, Danish Cancer Society Research Center, Strandboulevarden 49, 2100 Copenhagen, Denmark b Redox Signaling and Oxidative Stress Group, Cell Stress and Survival Unit, Danish Cancer Society Research Center, Strandboulevarden 49, 2100 Copenhagen, Denmark c Department of Biology, University of Rome Tor Vergata, Rome, Italy d Translational Disease Systems Biology, Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research University of Copenhagen, Copenhagen, Denmark article info abstract Article history: Nitric oxide (NO) is an essential signaling molecule in the regulation of multiple cellular processes. It is endoge- Received 21 December 2018 nously synthesized by NO synthase (NOS) as the product of L-arginine oxidation to L-citrulline, requiring NADPH, Received in revised form 17 March 2019 molecular oxygen, and a pterin cofactor.
    [Show full text]
  • Soluble Guanylate Cyclase and Cgmp-Dependent Protein Kinase I Expression in the Human Corpus Cavernosum
    International Journal of Impotence Research (2000) 12, 157±164 ß 2000 Macmillan Publishers Ltd All rights reserved 0955-9930/00 $15.00 www.nature.com/ijir Soluble guanylate cyclase and cGMP-dependent protein kinase I expression in the human corpus cavernosum T Klotz1*, W Bloch2, J Zimmermann1, P Ruth3, U Engelmann1 and K Addicks2 1Department of Urology, University of Cologne; 2Institute I of Anatomy, University of Cologne; and 3Institute of Pharmacology, TU University of Munich, Germany Nitric oxide (NO) as a mediator in smooth muscle cells causes rapid and robust increases in cGMP levels. The cGMP-dependent protein kinase I has emerged as an important signal transduction mediator for smooth muscle relaxation. The purpose of this study was to examine the existence and distribution of two key enzymes of the NO=cGMP pathway, the cGMP-dependent kinase I (cGK I) and the soluble guanylate cyclase (sGC) in human cavernosal tissue. The expression of the enzymes were examined in corpus cavernosum specimens of 23 patients. Eleven potent patients suffered from penile deviations and were treated via Nesbit's surgical method. Nine long-term impotent patients underwent implantation of ¯exible hydraulic prothesis. Three potent patients underwent trans-sexual operations. Expression of the sGC and cGK I were examined immunohistochemically using speci®c antibodies. In all specimens of cavernosal tissue a distinct immunoreactivity was observed in different parts and structures. We found a high expression of sGC and cGK I in smooth muscle cells of vessels and in the ®bromuscular stroma. The endothelium of the cavernosal sinus, of the cavernosal arteries, and the cavernosal nerve ®bers showed an immunoreactivity against sGC.
    [Show full text]
  • Safe Handling of Sodium Azide (SAZ)
    Safe Handling of Sodium Azide (SAZ) 1,2 Sodium azide (SAZ, CAS# 26628-22-8) is a white crystalline solid [molecular formula of (NaN3)] used in organic synthesis and also as a well-known preservative at low concentrations in molecular biology reagents. Azide chemistry3,4 offers an effective means to synthesize a range of nitrogen-containing compounds with a wide variety of functional groups. But SAZ poses some significant risks. It is highly toxic and can react to form potentially explosive compounds. Azide reagents and intermediates react with some metals5, strong acids, and certain chlorinated solvents6 and this needs to be considered when using SAZ or when developing routes that involve azide-containing intermediates. Health Hazards & Physical Hazards of SAZ Hazard Statements Fatal if swallowed or in contact with skin. Very toxic to aquatic life with long lasting effects. Health Hazards: SAZ is highly toxic when ingested orally or absorbed through the skin. Azides form strong complexes with hemoglobin, and consequently block oxygen transport in the blood. They are more harmful to the heart and the brain than to other organs, because the heart and the brain use a lot of oxygen. Symptoms of exposure include headache, dizziness, nausea and vomiting, rapid breathing and heart rate, and skin burns and blisters (direct skin contact) and in the case of serious overexposure, convulsions and death. It will also react with acids to form hydrazoic acid (HN3). Unlike sodium azide, which is a crystalline solid, hydrazoic acid is a low-boiling, volatile, liquid. Hydrazoic acid is also highly toxic and its volatility makes it more readily inhaled causing lung irritation and potentially bronchitis and lung edema.
    [Show full text]
  • Effects of Tetrahydrobiopterin on Endothelial Dysfunction in Rats with Ischemic Acute Renal Failure
    J Am Soc Nephrol 11: 301–309, 2000 Effects of Tetrahydrobiopterin on Endothelial Dysfunction in Rats with Ischemic Acute Renal Failure MASAO KAKOKI,* YASUNOBU HIRATA,* HIROSHI HAYAKAWA,* ETSU SUZUKI,* DAISUKE NAGATA,* AKIHIRO TOJO,* HIROAKI NISHIMATSU,* NOBUO NAKANISHI,‡ YOSHIYUKI HATTORI,§ KAZUYA KIKUCHI,† TETSUO NAGANO,† and MASAO OMATA* *The Second Department of Internal Medicine, Faculty of Medicine, and †Faculty of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan; ‡Department of Biochemistry, Meikai University School of Dentistry, Saitama, Japan; and §Department of Endocrinology, Dokkyo University School of Medicine, Tochigi, Japan. Abstract. The role of nitric oxide (NO) in ischemic renal injury 4 fmol/min per g kidney; serum creatinine: control 23 Ϯ 2, is still controversial. NO release was measured in rat kidneys ischemia 150 Ϯ 27, ischemia ϩ BH4 48 Ϯ 6 ␮M; mean Ϯ subjected to ischemia and reperfusion to determine whether SEM). Most of renal NOS activity was calcium-dependent, and (6R)-5,6,7,8-tetrahydro-L-biopterin (BH4), a cofactor of NO its activity decreased in the ischemic kidney. However, it was synthase (NOS), reduces ischemic injury. Twenty-four hours restored by BH4 (control 5.0 Ϯ 0.9, ischemia 2.2 Ϯ 0.4, after bilateral renal arterial clamp for 45 min, acetylcholine- ischemia ϩ BH4 4.3 Ϯ 1.2 pmol/min per mg protein). Immu- induced vasorelaxation and NO release were reduced and renal noblot after low-temperature sodium dodecyl sulfate-polyac- excretory function was impaired in Wistar rats. Administration rylamide gel electrophoresis revealed that the dimeric form of of BH4 (20 mg/kg, by mouth) before clamping resulted in a endothelial NOS decreased in the ischemic kidney and that it marked improvement of those parameters (10Ϫ8 M acetylcho- was restored by BH4.
    [Show full text]
  • Looking for New Classes of Bronchodilators
    REVIEW BRONCHODILATORS The future of bronchodilation: looking for new classes of bronchodilators Mario Cazzola1, Paola Rogliani 1 and Maria Gabriella Matera2 Affiliations: 1Dept of Experimental Medicine, University of Rome Tor Vergata, Rome, Italy. 2Dept of Experimental Medicine, University of Campania “Luigi Vanvitelli”, Naples, Italy. Correspondence: Mario Cazzola, Dept of Experimental Medicine, University of Rome Tor Vergata, Via Montpellier 1, Rome, 00133, Italy. E-mail: [email protected] @ERSpublications There is a real interest among researchers and the pharmaceutical industry in developing novel bronchodilators. There are several new opportunities; however, they are mostly in a preclinical phase. They could better optimise bronchodilation. http://bit.ly/2lW1q39 Cite this article as: Cazzola M, Rogliani P, Matera MG. The future of bronchodilation: looking for new classes of bronchodilators. Eur Respir Rev 2019; 28: 190095 [https://doi.org/10.1183/16000617.0095-2019]. ABSTRACT Available bronchodilators can satisfy many of the needs of patients suffering from airway disorders, but they often do not relieve symptoms and their long-term use raises safety concerns. Therefore, there is interest in developing new classes that could help to overcome the limits that characterise the existing classes. At least nine potential new classes of bronchodilators have been identified: 1) selective phosphodiesterase inhibitors; 2) bitter-taste receptor agonists; 3) E-prostanoid receptor 4 agonists; 4) Rho kinase inhibitors; 5) calcilytics; 6) agonists of peroxisome proliferator-activated receptor-γ; 7) agonists of relaxin receptor 1; 8) soluble guanylyl cyclase activators; and 9) pepducins. They are under consideration, but they are mostly in a preclinical phase and, consequently, we still do not know which classes will actually be developed for clinical use and whether it will be proven that a possible clinical benefit outweighs the impact of any adverse effect.
    [Show full text]
  • Synthesis and Consecutive Reactions of Α-Azido Ketones: a Review
    Molecules 2015, 20, 14699-14745; doi:10.3390/molecules200814699 OPEN ACCESS molecules ISSN 1420-3049 www.mdpi.com/journal/molecules Review Synthesis and Consecutive Reactions of α-Azido Ketones: A Review Sadia Faiz 1,†, Ameer Fawad Zahoor 1,*, Nasir Rasool 1,†, Muhammad Yousaf 1,†, Asim Mansha 1,†, Muhammad Zia-Ul-Haq 2,† and Hawa Z. E. Jaafar 3,* 1 Department of Chemistry, Government College University Faisalabad, Faisalabad-38000, Pakistan, E-Mails: [email protected] (S.F.); [email protected] (N.R.); [email protected] (M.Y.); [email protected] (A.M.) 2 Office of Research, Innovation and Commercialization, Lahore College for Women University, Lahore-54600, Pakistan; E-Mail: [email protected] 3 Department of Crop Science, Faculty of Agriculture, Universiti Putra Malaysia, Serdang-43400, Selangor, Malaysia † These authors contributed equally to this work. * Authors to whom correspondence should be addressed; E-Mails: [email protected] (A.F.Z.); [email protected] (H.Z.E.J.); Tel.: +92-333-6729186 (A.F.Z.); Fax: +92-41-9201032 (A.F.Z.). Academic Editors: Richard A. Bunce, Philippe Belmont and Wim Dehaen Received: 20 April 2015 / Accepted: 3 June 2015 / Published: 13 August 2015 Abstract: This review paper covers the major synthetic approaches attempted towards the synthesis of α-azido ketones, as well as the synthetic applications/consecutive reactions of α-azido ketones. Keywords: α-azido ketones; synthetic applications; heterocycles; click reactions; drugs; azides 1. Introduction α-Azido ketones are very versatile and valuable synthetic intermediates, known for their wide variety of applications, such as in amine, imine, oxazole, pyrazole, triazole, pyrimidine, pyrazine, and amide alkaloid formation, etc.
    [Show full text]
  • Structural Perspectives on the Mechanism of Soluble Guanylate Cyclase Activation
    International Journal of Molecular Sciences Review Structural Perspectives on the Mechanism of Soluble Guanylate Cyclase Activation Elizabeth C. Wittenborn and Michael A. Marletta * California Institute for Quantitative Biosciences, Departments of Chemistry and of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA; [email protected] * Correspondence: [email protected] Abstract: The enzyme soluble guanylate cyclase (sGC) is the prototypical nitric oxide (NO) receptor in humans and other higher eukaryotes and is responsible for transducing the initial NO signal to the secondary messenger cyclic guanosine monophosphate (cGMP). Generation of cGMP in turn leads to diverse physiological effects in the cardiopulmonary, vascular, and neurological systems. Given these important downstream effects, sGC has been biochemically characterized in great detail in the four decades since its discovery. Structures of full-length sGC, however, have proven elusive until very recently. In 2019, advances in single particle cryo–electron microscopy (cryo-EM) enabled visualization of full-length sGC for the first time. This review will summarize insights revealed by the structures of sGC in the unactivated and activated states and discuss their implications in the mechanism of sGC activation. Keywords: nitric oxide; soluble guanylate cyclase; cryo–electron microscopy; enzyme structure Citation: Wittenborn, E.C.; Marletta, 1. Introduction M.A. Structural Perspectives on the Soluble guanylate cyclase (sGC) is a nitric oxide (NO)-responsive enzyme that serves Mechanism of Soluble Guanylate as a source of the secondary messenger cyclic guanosine monophosphate (cGMP) in Cyclase Activation. Int. J. Mol. Sci. humans and other higher eukaryotes [1]. Upon NO binding to sGC, the rate of cGMP 2021, 22, 5439.
    [Show full text]
  • Nitric Oxide Activates Guanylate Cyclase and Increases Guanosine 3':5'
    Proc. Natl. Acad. Sci. USA Vol. 74, No. 8, pp. 3203-3207, August 1977 Biochemistry Nitric oxide activates guanylate cyclase and increases guanosine 3':5'-cyclic monophosphate levels in various tissue preparations (nitro compounds/adenosine 3':5'-cyclic monophosphate/sodium nitroprusside/sodium azide/nitrogen oxides) WILLIAM P. ARNOLD, CHANDRA K. MITTAL, SHOJI KATSUKI, AND FERID MURAD Division of Clinical Pharmacology, Departments of Medicine, Pharmacology, and Anesthesiology, University of Virginia, Charlottesville, Virginia 22903 Communicated by Alfred Gilman, May 16, 1977 ABSTRACT Nitric oxide gas (NO) increased guanylate cy- tigation of this activation. NO activated all crude and partially clase [GTP pyrophosphate-yase (cyclizing), EC 4.6.1.21 activity purified guanylate cyclase preparations examined. It also in- in soluble and particulate preparations from various tissues. The effect was dose-dependent and was observed with all tissue creased cyclic GMP but not adenosine 3':5'-cyclic monophos- preparations examined. The extent of activation was variable phate (cyclic AMP) levels in incubations of minces from various among different tissue preparations and was greatest (19- to rat tissues. 33-fold) with supernatant fractions of homogenates from liver, lung, tracheal smooth muscle, heart, kidney, cerebral cortex, and MATERIALS AND METHODS cerebellum. Smaller effects (5- to 14-fold) were observed with supernatant fractions from skeletal muscle, spleen, intestinal Male Sprague-Dawley rats weighing 150-250 g were decapi- muscle, adrenal, and epididymal fat. Activation was also ob- tated. Tissues were rapidly removed, placed in cold 0.-25 M served with partially purified preparations of guanylate cyclase. sucrose/10 mM Tris-HCl buffer (pH 7.6), and homogenized Activation of rat liver supernatant preparations was augmented in nine volumes of this solution by using a glass homogenizer slightly with reducing agents, decreased with some oxidizing and Teflon pestle at 2-4°.
    [Show full text]
  • Fabricating Water Dispersible Superparamagnetic Iron Oxide Nanoparticles for Biomedical Applications Through Ligand Exchange and Direct Conjugation
    Nanomaterials 2016, 6, 100; doi:nano6060100 S1 of S10 Supplementary Materials: Fabricating Water Dispersible Superparamagnetic Iron Oxide Nanoparticles for Biomedical Applications through Ligand Exchange and Direct Conjugation Tina Lam, Pramod K. Avti, Philippe Pouliot, Foued Maafi, Jean-Claude Tardif, Éric Rhéaume, Frédéric Lesage and Ashok Kakkar Experimental The following compounds were purchased and used as received; tetraethylene glycol, tetraethylene glycol monomethyl ether, p-toluene sulfonyl chloride (tosyl chloride), methane sulfonyl chloride (mesyl chloride), sodium hydroxide, lithium aluminium hydride (LiAlH4), 4- dimethylaminopyridine (DMAP), triethylamine, diethyl amine, sodium azide, tetrabutylammonium iodide, sodium ascorbate (Na Ascorbate), copper(II) sulphate hexahydrate, borane-THF, bis(triphenylphosphine) palladium(II) dichloride, 1-bromo-4-iodo benzene, disodium ethylenediamine tetraacetate (EDTA), iron(II) chloride tetrahydrate (FeCl2·4H2O), iron (III) chloride hexahydrate (FeCl3·6H2O), and oleic acid from Sigma Aldrich (St. Louis, MO, USA), triisopropylsilyl acetylene and trimethylsilyl acetylene from Oakwoods Chemicals (Estill, SC, USA), 1-ethyl-3-(3- dimethylaminopropyl) carbodiimide from Chem Impex International (Wood Dale, IL, USA), and Iron standard # 13830 from Alfa Aesar (Haverhill, MA, USA). Polyvinylidene Fluoride (PVDF) Syringe filters of 17 mm diameter and pore size 0.45 μm were purchased from Sterlitech (Kent, WA, USA). The solvents triethylamine, diethylamine, tetrahydrofuran, acetic acid, acetic anhydride, acetone, chloroform, dichloromethane, benzene, methanol and toluene were purchased from Fisher scientific and ACP Chemicals and used as received. Solvents were obtained from drying columns and purged under N2 prior to use. Milli-Q Ultrapure water was doubly distilled by reverse osmosis though a Millipore RiOS8, followed by filtration through a Milli-Q Academic A10 filtration unit prior to use.
    [Show full text]