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Molecular Identification and Expression Analysis of Carbonic Anhydrase VII in Pufferfish (Takifugu rubripes)

  • Kanij Rukshana Sumi
  • Soo Cheol Kim
  • Jewel Howlader
  • Md Rajib Sharker
  • Kap Seong Choi
  • Sang Ki Choi
  • Jong-In Park
  • Ill-Sup Nou
  • Kang Hee KhoEmail author
Article
  • 8 Downloads

Abstract

A carbonic anhydrase VII gene, encoding 277 amino acids, was identified in the intestinal tissue of pufferfish (Takifugu rubripes). The translated protein with an 833-bp complete coding sequence derived from the 1378-bp cloned sequence showed 83% identity with swamp eel CA VII, 76% with zebrafish CA VII, and 77% with coho salmon CA VII-like protein. The cloned protein also showed 68–69% identity with mammalian CA VII. The predicted molecular weight and iso-electric point of the protein were 30.84 kDa and 6.07, respectively. Active site analysis of the pufferfish CA VII indicated that most of the important residues involved in catalytic activity were highly conserved, whereas four cysteine residues at positions 55, 103, 184, and 275 differed from those in human CA II, and were related to cell-defense mechanisms against oxidative damage. Phylogenetic analysis showed that the cloned sequence was clustered within the fish CA VII clade and close to the swamp eel CA VII. Structural modeling of the pufferfish CA VII protein revealed the conservation of zinc binding histidine residues (zinc ion and histidine residues). Differential expression patterns of the pufferfish CA VII were determined with semiquantitative reverse transcription (RT)-PCR and quantitative PCR (q-PCR) as well. The results of q-PCR revealed that the pufferfish CA VII was highly expressed in intestinal tissue. The pufferfish CA VII was also detected within intestinal tissue sections using an in situ hybridization assay.

Keywords

pufferfish Takifugu rubripes carbonic anhydrase (CA) VII intestinal tissue 

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Notes

Acknowledgments

This paper was supported by Jeonnam Sea Grant R&D program of 2018 [Grant no. 20180330]. Thanks are extended to Chungnam Prefectural Fisheries Institute for providing experimental fishes.

References

  1. Ahmed YA, El-Hafez AAE, Zayed AE (2009) Histological and histochemical studies on the esophagus, stomach and small intestines of Varanus niloticus. J Vet Anat 2:35–48CrossRefGoogle Scholar
  2. Alterio V, Di Fiore A, D’Ambrosio KATIA, Supuran CT, De Simone G (2009) X-ray crystallography of carbonic anhydrase inhibitors and its importance in drug design. In: Supuran CT, Winum JY (eds) Drug design of zinc-enzyme inhibitors: functional, structural, and disease applications. Wiley & Sons, Hoboken, pp 73–138Google Scholar
  3. Alva V, Nam SZ, Söding J, Lupas AN (2016) The MPI bioinformatics toolkit as an integrative platform for advanced protein sequence and structure analysis. Nucleic Acids Res 44:W410–W415. doi: https://doi.org/10.1093/nar/gkw348 CrossRefGoogle Scholar
  4. Aspatwar A, Tolvanen MEE, Ortutay C, Parkkila S (2010) Carbonic anhydrase related protein VIII and its role in neurodegeneration and cancer. Curr Pharm Design 16:3264–3276CrossRefGoogle Scholar
  5. Berman HM, Westbrook J, Feng Z, Gillilan G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE (2000) The protein data bank. Nucleic Acids Res 28:235–242CrossRefGoogle Scholar
  6. Bootorabi F, Jänis J, Smith E, Waheed A, Kukkurainen S, Hytönen V, Valjakka J, Supuran CT, Vullo D, Sly WS, Parkkila S (2010) Analysis of a shortened form of human carbonic anhydrase VII expressed in vitro compared to the full-length enzyme. Biochimie 92:1072–1080CrossRefGoogle Scholar
  7. Bryman A, Cramer D (2009) Quantitative data analysis with SPSS 14, 15 & 16: a guide for social scientists. Taylor & Francis Ltd, New York, 408 pGoogle Scholar
  8. Del Giudice R, Monti DM, Truppo E, Arciello A, Supuran CT, De Simone G, Monti SM (2013) Human carbonic anhydrase VII protects cells from oxidative damage. Biol Chem 394:1343–1348CrossRefGoogle Scholar
  9. Di Fiore A, Truppo E, Supuran CT, Alterio V, Dathan N, Bootorabi F, Parkkila S, Monti SM, De Simone G (2010) Crystal structure of the C183S/C217S mutant of human CA VII in complex with acetazolamide. Bioorg Med Chem Lett 20:5023–5026CrossRefGoogle Scholar
  10. Eisenberg D, Lüthy R, Bowie JU (1997) VERIFY3D: assessment of protein models with three-dimensional profiles. Method Enzymol 277:396–404CrossRefGoogle Scholar
  11. Esbaugh AJ, Lund SG, Tufts BL (2004) Comparative physiology and molecular analysis of carbonic anhydrase from the red blood cells of teleost fish. J Comp Physiol B 174:429–438Google Scholar
  12. Esbaugh AJ, Perry SF, Bayaa M, Georgalis T, Nickerson J, Tufts BL, Gilmour KM (2005) Cytoplasmic carbonic anhydrase isozymes in rainbow trout Oncorhynchus mykiss: comparative physiology and molecular evolution. J Exp Biol 208:1951–1961CrossRefGoogle Scholar
  13. Esbaugh AJ, Tufts BL (2006a) Tribute to RG Boutilier: evidence of a high activity carbonic anhydrase isozyme in the red blood cells of an ancient vertebrate, the sea lamprey Petromyzon marinus. J Exp Biol 209:1169–1178CrossRefGoogle Scholar
  14. Esbaugh AJ, Tufts BL (2006b) The structure and function of carbonic anhydrase isozymes in the respiratory system of vertebrates. Resp Physiol Neurobi 154:185–198CrossRefGoogle Scholar
  15. Forster RP, Goldstein L, Rosen JK (1972) Intrarenal control of urea reabsorption by renal tubules of the marine elasmobranch, Squalus acanthias. Comp Biochem Phys A 42:3–12CrossRefGoogle Scholar
  16. Güzel Ö, Innocenti A, Scozzafava A, Salman A, Supuran CT (2009) Carbonic anhydrase inhibitors. Phenacetyl-, pyridylacetyl- and thienylacetyl-substituted aromatic sulfonamides act as potent and selective isoform VII inhibitors. Bioorg Med Chem Lett 19:3170–3173Google Scholar
  17. Hilvo M, Innocenti A, Monti SM, Simone GD, Supuran CT, Parkkila S (2008) Recent advances in research on the most novel carbonic anhydrases, CA XIII and XV. Curr Pharm Design 14:672–678CrossRefGoogle Scholar
  18. Lindskog S (1982) Carbonic anhydrase. Adv Inorg Biochem 4:115–170Google Scholar
  19. Lindskog S, Silverman D (2000) The catalytic mechanism of mammalian carbonic anhydrases. In: Chegwidden WR, Carter ND, Edwards YH (eds) The carbonic anhydrases. New Horizons, Basel, pp 175–195CrossRefGoogle Scholar
  20. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔct method. Methods 25:402–408CrossRefGoogle Scholar
  21. Lund SG, Dyment P, Gervais MR, Moyes CD, Tufts BL (2002) Characterization of erythrocyte carbonic anhydrase in an ancient fish, the longnose gar (Lepistosteus osseus). J Comp Physiol B 172:467–476CrossRefGoogle Scholar
  22. Montgomery JC, Venta PJ, Eddy RL, Fukushima YS, Shows TB, Tashian RE (1991) Characterization of the human gene for a newly discovered carbonic anhydrase, CA VII, and its localization to chromosome 16. Genomics 11:835–848CrossRefGoogle Scholar
  23. Rivera C, Voipio J, Kaila K (2005) Two developmental switches in GABAergic signalling: the K+-Cl cotransporter KCC2 and carbonic anhydrase CAVII. J Physiol 562:27–36CrossRefGoogle Scholar
  24. Ruusuvuori E, Li H, Huttu K, Palva JM, Smirnov S, Rivera C, Kaila K, Voipio J (2004) Carbonic anhydrase isoform VII acts as a molecular switch in the development of synchronous gamma-frequency firing of hippocampal CA1 pyramidal cells. J Neurosci 24:2699–2707CrossRefGoogle Scholar
  25. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–125Google Scholar
  26. Sali A, Blundell TL (1993) Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol 234:779–815CrossRefGoogle Scholar
  27. Santovito G, Marino SM, Sattin G, Cappellini R, Bubacco L, Beltramini M (2012) Cloning and characterization of cytoplasmic carbonic anhydrase from gills of four Antarctic fish: insights into the evolution of fish carbonic anhydrase and cold adaptation. Polar Biol 35:1587–1600CrossRefGoogle Scholar
  28. Sattin G, Mager EM, Beltramini M, Grosell M (2010) Cytosolic carbonic anhydrase in the Gulf toadfish is important for tolerance to hypersalinity. Comp Biochem Phys A 156:169–175CrossRefGoogle Scholar
  29. Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Söding J, Thompson JD, Higgins DG (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 7:539–544CrossRefGoogle Scholar
  30. Silverman DA, Vincent SH (1983) Proton transfer in the catalytic mechanism of carbonic anhydrase. CRC Cr Rev Bioch Mol 14:207–255CrossRefGoogle Scholar
  31. Sumi KR, Kim SC, Natarajan S, Choi KS, Choi MR, Kim HT, Park JI, Nou IS, Gilmour KM, Kho KH (2017) Molecular cloning and characterization of secretory carbonic anhydrase VI in pufferfish (Takifugu rubripes). Gene 640:57–65CrossRefGoogle Scholar
  32. Sumi KR, Nou IS, Kho KH (2016) Identification and expression of a novel carbonic anhydrase isozyme in the pufferfish Takifugu vermicularis. Gene 588:173–179CrossRefGoogle Scholar
  33. Supuran CT (2008) Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nat Rev Drug Discov 7:168–181CrossRefGoogle Scholar
  34. Supuran CT (2011) Carbonic anhydrase inhibitors and activators for novel therapeutic applications. Future Med Chem 3:1165–1180CrossRefGoogle Scholar
  35. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729CrossRefGoogle Scholar
  36. Tashian RE (1992) Genetics of the mammalian carbonic anhydrases. Adv Genet 30:321–349CrossRefGoogle Scholar
  37. Tashian RE, Hewett-Emmett D, Carter N, Bergenhem NC (2000) Carbonic anhydrase (CA)-related proteins (CA-RPs), and transmembrane proteins with CA or CA-RP domains. EXS 90:105–120Google Scholar
  38. Truppo E, Supuran CT, Sandomenico A, Vullo D, Innocenti A, Di Fiore A, Monti SM (2012) Carbonic anhydrase VII is S-glutathionylated without loss of catalytic activity and affinity for sulfonamide inhibitors. Bioorg Med Chem Lett 22:1560–1564CrossRefGoogle Scholar
  39. Tufts BL, Esbaugh A, Lund SG (2003) Comparative physiology and molecular evolution of carbonic anhydrase in the erythrocytes of early vertebrates. Comp Biochem Phys A 136:259–269CrossRefGoogle Scholar
  40. Ugochukwu E, Shafqat N, Pilka E, Chaikuad A, Krojer T, Muniz J, Kim J, Bray J, Bountra C, Arrowsmith CH, Weigelt J, Edwards A, von Delft F, Carpenter EP, Yue WW, Oppermann U (2010) 3MDZ, crystal structure of human carbonic anhydrase VII [isoform 1], CA7, RCSB PDB. http://www.rcsb.org/structure/3MDZ Accessed 4 Mar 2019
  41. Waterhouse AM, Procter JB, Martin DMA, Clamp M, Barton GJ (2009) Jalview Version 2 — a multiple sequence alignment editor and analysis workbench. Bioinformatics 25:1189–1191CrossRefGoogle Scholar

Copyright information

© KSO, KIOST and Springer 2019

Authors and Affiliations

  • Kanij Rukshana Sumi
    • 1
    • 2
  • Soo Cheol Kim
    • 1
  • Jewel Howlader
    • 3
    • 4
  • Md Rajib Sharker
    • 1
  • Kap Seong Choi
    • 5
  • Sang Ki Choi
    • 6
  • Jong-In Park
    • 3
  • Ill-Sup Nou
    • 3
  • Kang Hee Kho
    • 1
    Email author
  1. 1.Department of Fisheries Science, College of Fisheries and Ocean SciencesChonnam National UniversityYeosuKorea
  2. 2.Department of AquaculturePatuakhali Science and Technology UniversityDhakaBangladesh
  3. 3.Department of Horticulture, College of Life Science and Natural ResourcesSunchon National UniversitySuncheonKorea
  4. 4.Department of HorticulturePatuakhali Science and Technology UniversityDhakaBangladesh
  5. 5.Department of Food Science, College of Life Science and Natural ResourcesSunchon National UniversitySuncheonKorea
  6. 6.Department of Biology, College of Life Science and Natural ResourcesSunchon National UniversitySuncheonKorea

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