İnsan Eritrositlerinden Saflaştırılan Karbonik Anhidraz (CA) I ve II İzoenzimleri Üzerinde Eozin Y'nin İnhibisyon Etkisi

Yıl 2020, Cilt: 10 Sayı: 1, 338 - 344, 01.03.2020

Zuhal Alım

https://doi.org/10.21597/jist.593758

Cited By: 2

Öz

Bütün hücrelerde metabolizma sonucu açığa çıkan ve vücuttan atılması gereken karbondioksit üretilir. Bu CO2'nin büyük bir kısmı eritrositlerde CA enzimi tarafından bikarbonata dönüştürülür ve vücuttan atılır. Bu nedenle, CA'nın kırmızı kan hücrelerinde hayati bir rolü vardır. Ayrıca, CA diğer birçok patolojik ve fizyolojik süreçlere dahil olmuştur ve CA inhibitörlerinin, özellikle glokom gibi birçok hastalığın tedavisinde ve tanısında etkili olduğu belirlenmiştir. CA inhibitörlerinin önemi göz önüne alındığında, bu çalışmada Eosin Y'nin insan eritrositlerinden saflaştırılmış CA I ve CA II izoenzimlerin aktivitesi üzerindeki inhibisyon etkilerinin araştırılması amaçlanmıştır. Eozin Y, histolojik ve tıbbi uygulamalarda yaygın olarak kullanılan bir boya molekülüdür. Bu amaçla öncelikle CA I ve CA II izoenzimleri, Sepharose-4B-L-tirozin-sülfanilamid afinite kromatografısi kullanılarak insan eritrositlerinden saflaştırıldı. Daha sonra, Eozin Y'nin bu insan eritrosit CA I (hCA I) ve CA II (hCA II) izoenzimlerinin aktivitesi üzerindeki inhibitör etkisi araştırıldı. hCA I ve hCA II'nin millimolar aralıktaki Eozin Y tarafından inhibe edildiği belirlendi. IC50 değerleri hCA I için 3,78 mM ve hCA II için 2,04 mM. Ki değerleri ise hCA I ve hCA II için sırasıyla 9,65 ± 0,968 mM ve 7,52 ± 2,88 mM olarak tespit edildi. Sonuç olarak, bu çalışmada elde edilen sonuçların, ilaç adayı olabilecek yeni CA inhibitörlerinin geliştirilmesinde faydalı olacağı ümit edilmektedir.

Anahtar Kelimeler

İnsan eritrositi, karbonik anhidraz, eozin Y, inhibisyon

Inhibition Effect of Eosin Y on Carbonic Anhydrase (CA) I and II Isoenzymes Purified from Human Erythrocytes

Öz

All cells produce carbon dioxide (CO2), which is released as a result of metabolism and must be removed from the body. A large part of this CO2 is converted to bicarbonate by the carbonic anhydrase (CA) enzyme in erythrocytes and is discarded from the body. So, CA has a vital role in red blood cells. In addition to, CA involved in many other pathological and physiological processes and it was determined that the inhibitors of CA were effective in the treatment and diagnosis of many diseases particularly glaucoma. Considering the importance of the CA's inhibitors, in this study it was intended to research the inhibition effects of Eosin Y on CA I and CA II isoenzymes activity purified from human erythrocytes. Eosin Y is a dye molecule commonly used in histological and medical applications. For this purpose, firstly CA I and CA II isoenzymes were purified from human erythrocytes by using Sepharose-4B-L-tyrosine-sulfanilamide affinity chromatography. Then the inhibitory effect of Eosin Y on the activity of these human erythrocyte CA I (hCA I) and CA II (hCA II) isoenzymes was investigated. It was determined that hCA I and hCA II were inhibited by Eosin Y in the millimolar range. IC50 values were found to be 3.78 mM for hCA I and 2.04 mM for hCA II and Ki values were determined as 9.65±0.968 mM and 7.52±2.88 mM for hCA I and hCA II, respectively. In conclusion, it is hoped that the results obtained in this study may be beneficial in the development of new CA inhibitors which may be drug candidates.

Anahtar Kelimeler

Human erythrocyte, carbonic anhydrase, eosin Y, inhibition

Kaynakça

• Aggarwal M, Kondeti B, McKenna R, 2013. Anticonvulsant/antiepileptic carbonic anhydrase inhibitors: a patent review. Expert Opinion on Therapeutic Patents, 23 (6): 717-724.
• Aksu K, Nar M, Tanç M, Vullo D, Gulcin I, Göksu F, Tümer F, Supuran CT, 2013. The synthesis of sulfamide analogues of dopamine related compounds and their carbonic anhydrase inhibitory properties. Biorganic Medicinal Chemistry, 21 (11): 2925-2931.
• Alım Z, Kilinc N, Sengul B, Beydemir S, 2015. Some Anti-Inflammatory Agents Inhibit Esterase Activities of Human Carbonic Anhydrase Isoforms i and II: An in Vitro Study. Chemical Biology and Drug Design, 86 (4): 857-863.
• Alım Z, 2018. 1H-indazole molecules reduced the activity of human erythrocytes carbonic anhydrase I and II isoenzymes. Journal of Biochemical and Molecular Toxicology, 32 (9): e22194. Alterio V, Di Fiore A, D'Ambrosio K, Supuran CT, De Simone G, 2012. Multiple binding modes of inhibitors to carbonic anhydrases: how to design specific drugs targeting 15 different isoforms. Chemical Reviews, 112 (8): 4421-4468.
• Aslan HE, Demir Y, Özaslan MS, Türkan F, Beydemir Ş, Küfrevioğlu ÖI. 2019. The behavior of some chalcones on acetylcholinesterase and carbonic anhydrase activity. Drug and Chemical Toxicology, 42(6):634-640.
• Benej M, Pastorekova S, Pastorek J, 2014. Carbonic anhydrase IX: regulation and role in cancer. Subcellular Biochemistry, 75: 199-219.
• Bradford MM, 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72 (1-2): 248–251.
• Buzas GM, Supuran CT, 2016. The history and rationale of using carbonic anhydrase inhibitors in the treatment of peptic ulcers. In memoriam Ioan Puşcaş (1932-2015). Journal of Enzyme Inhibition Medicinal Chemistry, 31 (4): 527-533.
• Caglayan C, Taslimi P, Demir Y, Küçükler S, Kandemir, FM, Gülcin I. 2019. The effects of zingerone against vancomycin‐induced lung, liver, kidney and testis toxicity in rats: The behavior of some metabolic enzymes. Journal of Biochemical and Molecular Toxicology, e22381. https://doi.org/10.1002/jbt.22381
• Caglayan C, Demir Y, Küçükler S, Taslimi P, Kandemir, FM, Gülcin I. 2018. The effects of hesperidin on sodium arsenite‐induced different organ toxicity in rats on metabolic enzymes as antidiabetic and anticholinergics potentials: A biochemical approach. Journal of Food Biochemistry, 43(2): e 12720.
• Chiaramonte N, Romanelli MN, Teodori E, Supuran CT, 2018. Amino Acids as Building Blocks for Carbonic Anhydrase Inhibitors. Metabolites, 8(2): 36.
• Ekinci D, Beydemir S, Alim Z, 2007. Some drugs inhibit in vitro hydratase and esterase activities of human carbonic anhydrase-I and II. Pharmacological Reports, 59(5): 580-587.
• Erdemir F, Celepci DB, Aktaş A, Gök Y, Kaya R, Taslimi P, Demir Y, Gulçin İ. 2019. Novel 2-aminopyridine liganded Pd(II) N-heterocyclic carbene complexes: Synthesis, characterization, crystal structure and bioactivity properties. Bioorganic Chemistry, 91:103134.
• Deitmer JW, Theparambil SM, Ruminot I, Becker HM, 2014. The role of membrane acid/base transporters and carbonic anhydrases for cellular pH and metabolic processes. Front Neuroscience, 8: 430.
• Demir Y, Oruç E, Topal A. 2016. Carbonic anhydrase activity responses and histopathological changes in gill and liver tissues after acute exposure to chromium in brown trout juveniles. Hacettepe Journal of Biology and Chemistry 44:515-523.
• Hassan I, Shajee B, Waheed A, Ahmad F, Sly WS, 2013. Structure, function and applications of carbonic anhydrase isozymes. Bioorganic & Medicinal Chemistry, 21 (6), 1570–1582.
• Hu PY, Roth DE, Skaggs LA, Venta PJ, Tashian RE, Guibaud P, Sly WS, 1992. A splice junction mutation in intron 2 of the carbonic anhydrase II gene of osteopetrosis patients from Arabic countries. Human Mutation, 1(4): 288-292.
• Jang BG, Yun SM, Ahn K, Song JH, Jo SA, Kim YY, Kim DK, Park MH, Han C, Koh YH, 2010. Plasma carbonic anhydrase II protein is elevated in Alzheimer's disease. Journal of Alzheimer's Disease, 21 (3): 939-945.
• Juncal VR, Abdo JF, Paranhos AJ, Santos PT, 2015. Carbonic anhydrase inhibitors as fourth drug in primary glaucomas: Is it worth it? Canadian Journal of Ophthalmology, 50(4): 297-301.
• Laemmli UK, 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227 (5259): 680–685.
• Kaya ED, Ergun B, Demir Y, Alım Z, Beydemir S. 2019. The In Vitro Impacts of Some Plant Extracts on Carbonic Anhydrase I, II and Paraoxonase-1. Hacettepe Journal of Biology and Chemistry, 47(1):51-59.
• Lineweaver H, Burk D, 1934. The determination of enzyme dissociation constants. Journal of the American chemical society, 56 (3): 658–666.
• Mori S, Kou I, Sato H, Emi M, Ito H, Hosoi T, Ikeqawa S, 2009. Nucleotide variations in genes encoding carbonic anhydrase 8 and 10 associated with femoral bone mineral density in Japanese female with osteoporosis. Journal of Bone and Mineral Metabolism. 27 (2): 213-216.
• Wolfensberger TJ, 1999. The role of carbonic anhydrase inhibitors in the management of macular edema. Documenta Ophthalmologica, 97 (3-4): 387-397.
• Rahman H, 2017. Utilization of eosin dye as an ion pairing agent for determination of pharmaceuticals: A brief review. International Journal of Pharmacy and Pharmaceutical Sciences, 9 (12):1-9
• Sağlık BN, Çevik UA, Osmaniye D, Levent S, Çavuşoğlu BK, Demir Y, Ilgın S, Özkay Y, Koparal AS, Beydemir Ş, Kaplancıklı ZA. 2019. Synthesis, molecular docking analysis and carbonic anhydrase I-II inhibitory evaluation of new sulfonamide derivatives. Bioorganic Chemistry, 91:103153.
• Scozzafava A, Supuran CT, Carta F, 2013. Antiobesity carbonic anhydrase inhibitors: a literature and patent review. Expert Opinion on Therapeutic Patents, 23 (6): 725-735.
• Sun MK, Alkon DL, 2002. Carbonic anhydrase gating of attention: memory therapy and enhancement. Trends in Pharmacological Sciences, 23 (2): 83-89.
• Supuran CT, Briganti F, Tilli S, Chegwidden WR, Scozzafava A, 2001. Carbonic anhydrase inhibitors: sulfonamides as antitumor agents? Bioorganic Medicinal Chemistry, 9(3): 703-14.
• Supuran CT, 2008. Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nature Reviews Drug Discovery, 7(2): 168-181.
• Supuran CT, 2011. Carbonic anhydrase inhibitors and activators for novel therapeutic applications. Future Medicinal Chemistry, 3 (9):1165-1180.
• Supuran CT, 2018. Carbonic anhydrase activators. Future Medicinal Chemistry, 10(5):561-573.
• Taslimi P, Kandemir, FM, Demir Y, İleritürk M, Temel Y, Caglayan C, Gülcin I. 2019. The Antidiabetic and Anticholinergic Effects of Chrysin on Cyclophosphamide-Induced Multiple Organs Toxicity in Rats: Pharmacological Evaluation of Some Metabolic Enzymes Activities. Journal of Biochemical and Molecular Toxicology, 33(6):e22313.
• Topal A, Atamanalp M, Oruç E, Demir Y, Beydemir Ş, Işık A. 2014. In vivo changes in carbonic anhydrase activity and histopathology of gill and liver tissues after acute exposure to chlorpyrifos in rainbow trout. Archives of Industrial Hygiene and Toxicology, 65(4) 377-385.
• Türkeş C, Arslan M, Demir Y, Çoçaj L, Rifati Nixha A, Beydemir Ş. 2019. Synthesis, biological evaluation and in silico studies of novel N-substituted phthalazine sulfonamide compounds as potent carbonic anhydrase and acetylcholinesterase inhibitors. Bioorganic Chemistry, 89:103004.
• Vaughan-Jones RD, Spitzer KW, 2002. Role of bicarbonate in the regulation of intracellular pH in the mammalian ventricular myocyte. Biochemistry and Cell Biology, 80 (5): 579-596.
• Verpoorte JA, Mehta S, Edsall JT, 1967. Esterase activities of human carbonic anhydrases B and C. The Journal of Biological Chemistry, 242 (18): 4221–4229.
• Wilbur KM, Anderson NG, 1948. Electrometric and colorimetric determination of carbonic anhydrase. The Journal of Biological Chemistry, 176(1): 147–154.
• Waheed AA, Rao Sridhar K, Gupta PD, 2000. Mechanism of Dye Binding in the Protein Assay Using Eosin Dyes. Analytical Biochemistry, 287 (1): 73-79.