Journals / Hacettepe Journal of Biology and Chemistry / 2012 / Cilt: 40 - Sayı: 2

Purification of Ca Isoenzymes from Human Cancerous Colon Tissue and Inhibitory Effects of Some Analgesics on Enzyme Activity

İnsan Kanserli Kolon Dokusundan CA İzoenzimlerinin Saflaştırılması ve Enzim Aktivitesi Üzerine Bazı Analjezik İlaçların İnhibitör Etkisi

Pages
127–132
Publication Date
2012-04-01
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

IC50 values were found as 0.0077, 0.025, 0.011 and 0.04 mM for dexketoprofen, pethidine, phenyramidol and tramadol respectively.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Carbonic Anhydrase CA is an enzyme which is responsible for the hydration of carbon dioxide to carbonic acid and it also takes places in many biological processes in the living organisms.
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Key Findings & Data:In this study, CA isoenzymes CA II and CA IX together were purified 78.4 fold with a yield of 54.86 and specific activity of 106.67 by using Sepharose 4B-L-tyrosine sulfanilamide affinity chromatography.
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Conclusion & Impact:IC50 values were found as 0.0077, 0.025, 0.011 and 0.04 mM for dexketoprofen, pethidine, phenyramidol and tramadol respectively.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Saflaştırma · Kanser · Kolon

Abstract

Carbonic Anhydrase CA is an enzyme which is responsible for the hydration of carbon dioxide to carbonic acid and it also takes places in many biological processes in the living organisms. In this study, CA isoenzymes CA II and CA IX together were purified 78.4 fold with a yield of 54.86 and specific activity of 106.67 by using Sepharose 4B-L-tyrosine sulfanilamide affinity chromatography. In SDS-PAGE molecular weights of CA II and CA IX were calculated as 29 kDa and 56 kDa respectively. Besides inhibitory effects of some analgesics on purified total enzyme was investigated. IC50 values were found as 0.0077, 0.025, 0.011 and 0.04 mM for dexketoprofen, pethidine, phenyramidol and tramadol respectively.

Özet

K arbonik anhidraz CA karbon dioksitin karbonik aside hidrasyonundan sorumlu bir enzimdir ve aynı zamanda canlı organizmalarda birçok biyolojik proseslerde yer almaktadır. Bu çalışmada, CA izoenzimleri CA II ve CA IX birlikte sepharose 4B-L-tirozin sülfanilamid afinite kromatografisi kullanılarak 106.67 spesifik aktivite ile % 54.86 verimle 78.4 kat saflaştırıldı. SDS-PAGE’de CA II ve CA IX izoenzimlerin molekül ağırlıkları sırası ile 29 kDa ve 56 kDa olarak hesaplandı. Ayrıca saflaştırılan total enzim üzerine bazı analjezik ilaçların inhibisyon etkisi araştırıldı. Deksketoprofen, pethidin, pheniramidol, tramadol’ün IC50 değerleri sırasıyla 0.0077, 0.025, 0.011, ve 0.04 mM olarak bulundu

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Cite This Article

DEMİRDAĞ, R., YERLİKAYA, E., KÜFREVİOĞLU, Ö. İ., GÜNDOĞDU, C. (2012). Purification of Ca Isoenzymes from Human Cancerous Colon Tissue and Inhibitory Effects of Some Analgesics on Enzyme Activity. Hacettepe Journal of Biology and Chemistry, Cilt: 40 - Sayı: 2, 127–132. https://ajindex.com/articles/250185

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References (15)

  1. C.T. Supuran, A. Scozzafava, Carbonic Anhydrase Inhibitors, Current Medic. Chem., 1 ( 2001) 61
  2. J.B. Feldstein, D.N. Silverman, Purification and characterization of carbonicanhydrase from the saliva of the rat, J. Biol. Chem., 259 (1984) 5447
  3. S.R. Krungkrai, N. Suraveratum, S. Rochanakij, J. Krungkrai, Characterization of carbonic anhydrase in Plasmodium falciparum. International Journal of Parasitology, 31, (2001) 661
  4. N. Demir, Y. Demir, H. Nadaroglu, Carbonic anhydrase from bovine bone, Prep. Biochem. & Biotech., 31 (2001) 33
  5. S. Beydemir, M. Çiftçi, Ö. Küfrevioglu, Effects of gentamicine sulfate on enzyme activities of carbonic anhydrase from human erytrocytes in vitro and from rat erytrocytes in vivo, Biol. & Pharmac. Bull., 25 (2002) 966
  6. S. Beydemir, İ. Gülçin, Effects of melatonin on carbonic anhydrase from human erytrocyte in vitro and from rat erytrocytes in vivo, J. Enzyme Inhibition & Medic. Chem., 19 (2004) 193
  7. W.R. Chegwidden, S.J. Dodgson, I. M. Spencer, In the Carbonic Anhydrase-New Horizons, Birkhauser Verlag, Basel, ( 2000) 343
  8. W.R. Chegwidden, Y. Edwards, N. Carter, The Carbonic Anhydrases-New Horizons, Mol. Bases of Inher. Disease (Scriver, C. R., Beaudet, A. L.,Sly,W. S., and Valle, D., eds) 8th Ed., pp. (2000), 2165, McGraw-Hill, Inc.,NewYork
  9. C.T. Supuran, A. Scozzafava, Carbonic anhydrase inhibitors: Part 94. 1,3,4-thiadiazole-2-sulfonamide derivatives as antitumor agents?, Eur. J. Med. Chem, 35 (2000) 867
  10. C.T. Supuran, Indisulam: An anticancer sulfonamide in clinical development, Expert Opin Investig Drugs., 12 (2003) 283
  11. C.T. Supuran, A. Innocenti, A. Mastrolorenzo, A. Scozzafava, Antiviral sulfonamide derivatives, Mini. Rev. Med. Chem., 4 (2004) 189
  12. I. Nishimoiri, Carbonic anhydrase relatedproteins, in Carbonic Anhydrase, Its Inhibitors and ActiVators (Supuran, C. T., Scozzafava, A., and Conway, J., Eds.), Acatalytic CAs, CRC Press, Boca Raton, FL, (2004) 24
  13. M. Hilvo, M. Tolvanen, A. Clark, B.R. Shen, G.N. Shah, A. Waheed, P. Halmi, M. Hanninen, J.M. Hamalainen, M. Vihinen, WS. Sly, S. Parkkıla, Characterızatıon of CA XV, a new GPI-Anchored form of carbonıc anhydrase, Biochem. J., 392 (2005) 83, Part 1
  14. S.V. Ivanov, I. Kuzmin, M.H. Wei, S. Pack, B.E. Johnson, E.J. Stanbridge, M.I. Lerman, Down-regulation of transmembrane carbonic anhydrases in renal cell carcinoma cell lines by wild-type Von Hippel-Lindau transgenes, Pros. Natl. Acad. Sci., USA, 95 (1998) 12596
  15. J. Pastore