Isolation and sequencing of swine carbonic anhydrase VI, an enzyme expressed in the swine kidney

BMC Research Notes

Hideharu Ochiai¹  Kazuyoshi Arishima²  Nobutune Ichihara⁵  Kensuke Orito⁶  Shino Kamoshida⁷  Masaru Murakami⁷  Juro Yatsu⁴  Toshiho Nishita³ 

[1] Research Institute of Biosciences, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 252-5201, Japan;Laboratory of Anatomy II, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 252-5201, Japan;Laboratory of Physiology I, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 252-5201, Japan;Miyagi Prefectural Meat Sanitation Inspection Station, 314 Imaizumi, Sakuraoka, Yoneyamacho, Tome-city, Miyagi 987-0031, Japan;Laboratory of Anatomy I, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 252-5201, Japan;Laboratory of Physiology II, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 252-5201, Japan;Laboratory of Molecular Biology, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 252-5201, Japan

关键词: Kidney disease;    RT-PCR;    mRNA;    Swine kidney;    cDNA;    Carbonic anhydrase VI;   

Others  :  1134400
DOI  :  10.1186/1756-0500-7-116

 received in 2013-10-17, accepted in 2014-02-24,  发布年份 2014

【 摘 要 】

Background

Carbonic anhydrase VI (CA-VI) is produced by the salivary gland and is secreted into the saliva. Although CA-VI is found in the epithelial cells of distal straight tubule of swine kidneys, the exact function of CA-VI in the kidneys remains unclear.

Results

CA-VI was located in the epithelial cells of distal straight tubule of swine kidneys.

A full-length cDNA clone of CA-VI was generated from the swine parotid gland by reverse transcription polymerase chain reaction, using degenerate primers designed based on conserved regions of the same locus in human and bovine tissues.

The cDNA sequence was 1348 base pairs long and was predicted to encode a 317 amino acid polypeptide with a putative signal peptide of 17 amino acids. The deduced amino acid sequence of mature CA-VI was most similar (77.4%) to that of human CA-VI. CA-VI expression was confirmed in both normal and nephritic kidneys, as well as parotid. As the primers used in this study spanned two exons, the influence of genomic DNA was not detected. The expression of CA-VI was demonstrated in both normal and nephritic kidneys, and mRNA of CA-VI in the normal kidneys which was the normalised to an endogenous β–actin was 0.098 ± 0.047, while it was significantly lower in the diseased kidneys (0.012 ± 0.007). The level of CA-VI mRNA in normal kidneys was 19-fold lower than that of the parotid gland (1.887).

Conclusions

The localisation of CA-VI indicates that it may play a specialised role in the kidney.

【 授权许可】

   
2014 Nishita et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 3.

Figure 5.

【 参考文献 】

• [1]Hewett-Emmett D, Tashian RE: Functional diversity, conservation, and convergence in the evolution of the alpha-, beta-, and gamma-carbonic anhydrase gene families. Mol Phylogenet Evol 1996, 1:550-577.
• [2]Fujikawa-Adachi K, Nishimori I, Taguchi T, Onishi S: Human carbonic anhydrase XIV (CA14): cDNA cloning, mRNA expression, and mapping to chromosome 1. Genomics 1999, 61:74-81.
• [3]Lehtonen J, Shen B, Vihinen M, Casini A, Scozzafava A, Supuran CT, Parkkila AK, Saarnio J, Kivelä AJ, Waheed A, Sly WS, Parkkila S: Characterization of CA XIII, a novel member of the carbonic anhydrase isozyme family. J Biol Chem 2004, 279:2719-2727.
• [4]Carter MJ: Carbonic anhydrase: isoenzymes, properties, distribution, and functional significance. Biol Rev Camb Philos Soc 1972, 47:465-513.
• [5]Fernley RT, Coghlan JP, Wright RD: Purification and characterization of a high-Mr carbonic anhydrase from sheep parotid gland. Biochem J 1988, 249:201-207.
• [6]Murakami H, Sly WS: Purification and characterization of human salivary carbonic anhydrase. J Biol Chem 1987, 262:1382-1388.
• [7]Asari M, Miura K, Ichihara N, Nishita T, Amasaki H: Distribution of carbonic anhydrase isozyme VI in the developing bovine parotid gland. Cells Tissues Organs 2000, 167:18-24.
• [8]Nishita T, Sakomoto M, Ikeda T, Amasaki H, Shino M: Purification of carbonic anhydrase isozyme VI (CA-VI) from swine saliva. J Vet Med Sci 2001, 63:1147-1149.
• [9]Kasuya T, Shibata S, Kaseda M, Ichihara N, Nishita T, Murakami M, Asari M: Immunohistolocalization and gene expression of the secretory carbonic anhydrase isozymes (CA-VI) in canine oral mucosa, salivary glands and oesophagus. Anat Histol Embryol 2007, 36:53-57.
• [10]Parkkila S, Kaunisto K, Rajaniemi L, Kumpulainen T, Jokinen K, Rajaniemi H: Immunohistochemical localization of carbonic anhydrase isoenzymes VI, II, and I in human parotid and submandibular glands. J Histochem Cytochem 1990, 38:941-947.
• [11]Parkkila S, Parkkila AK, Lehtola J, Reinilä A, Södervik HJ, Rannisto M, Rajaniemi H: Salivary carbonic anhydrase protects gastroesophageal mucosa from acid injury. Dig Dis Sci 1997, 42:1013-1019.
• [12]Hooper LV, Beranek MC, Manzella SM, Baenziger JU: Differential expression of GalNAc-4-sulfotransferase and GalNAc-transferase results in distinct glycoforms of carbonic anhydrase VI in parotid and submaxillary glands. J Biol Chem 1995, 270:5985-5993.
• [13]Karhumaa P, Leinonen J, Parkkila S, Kaunisto K, Tapanainen J, Rajaniemi H: The identification of secreted carbonic anhydrase VI as a constitutive glycoprotein of human and rat milk. Proc Natl Acad Sci U S A 2001, 98:11604-11608.
• [14]Aldred P, Fu P, Barrett G, Penschow JD, Wright RD, Coghlan JP, Fernley RT: Human secreted carbonic anhydrase: cDNA cloning, nucleotide sequence, and hybridization histochemistry. Biochemistry 1991, 30:569-575.
• [15]Jiang W, Woitach JT, Gupta D: Sequence of bovine carbonic anhydrase VI: potential recognition sites for N-acetylgalactosaminyltransferase. Biochem J 1996, 318:291-296.
• [16]Sok J, Wang XZ, Batchvarova N, Kuroda M, Harding H, Ron D: CHOP-Dependent stress-inducible expression of a novel form of carbonic anhydrase VI. Mol Cell Biol 1999, 19:495-504.
• [17]Murakami M, Kasuya T, Matsuba C, Ichihara N, Nishita T, Fujitani H, Asari M: Nucleotide sequence and expression of a cDNA encoding canine carbonic anhydrase VI (CA-VI). DNA Seq 2003, 14:195-198.
• [18]Ochiai H, Kanemaki N, Kamoshida S, Murakami M, Ichihara N, Asari M, Nishita T: Determination of full-length cDNA nucleotide sequence of equine carbonic anhydrase VI and its expression in various tissues. J Vet Med Sci 2009, 71:1233-1237.
• [19]Nishita T, Itoh S, Arai S, Ichihara N, Arishima K: Measurement of carbonic anhydrase isozyme VI (CA-VI) in swine sera, colostrums, saliva, bile, seminal plasma and tissues. Anim Sci J 2011, 82:673-678.
• [20]Frohman MA, Dush MK, Martin GR: Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A 1988, 85:8998-9002.
• [21]Takara K, Yamamoto K, Matsubara M, Minegaki T, Takahashi M, Yokoyama T, Okumura K: Effects of α-adrenoceptor antagonists on ABCG2/BCRP-mediated resistance and transport. PLoS One 2012, 7(2):e30697.
• [22]von Heijne G: A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 1986, 14:4683-4690.
• [23]Fernley RT, Wright RD, Coghlan JP: Complete amino acid sequence of ovine salivary carbonic anhydrase. Biochemistry 1988, 27:2815-2820.
• [24]Fernley RT, Wright RD, Coghlan JP: Radioimmunoassay of carbonic anhydrase VI in saliva and sheep tissues. Biochem J 1991, 274:313-316.
• [25]Nishita T, Tanaka Y, Wada Y, Murakami M, Kasuya T, Ichihara N, Matsui K, Asari M: Measurement of carbonic anhydrase isozyme VI (CA-VI) in bovine sera, saliva, milk and tissues. Vet Res Commun 2007, 31:83-92.
• [26]Kitade K, Nishita T, Yamato M, Sakamoto K, Hagino A, Katoh K, Obara Y: Expression and localization of carbonic anhydrase in bovine mammary gland and secretion in milk. Comp Biochem Physiol Part A 2003, 134:349-354.
• [27]Purkerson JM, Schwartzm GJ: The role of carbonic anhydrases in renal physiology. Kidney Int 2007, 71:103-115.
• [28]Schwartz GJ: Physiology and molecular biology of renal carbonic anhydrase. J Nephrol 2002, 15:S61-S74.
• [29]Räisänen SR, Lehenkari P, Tasanen M, Rahkila P, Härkönen PL, Väänänen HK: Carbonic anhydrase III protects cells from hydrogen peroxide-induced apoptosis. FASEB J 1999, 13:513-522.
• [30]Gailly P, Jouret F, Martin D, Debaix H, Parreira KS, Nishita T, Blanchard A, Antignac C, Willnow TE, Courtoy PJ, Scheinman SJ, Christensen EI, Devuyst O: A novel renal carbonic anhydrase type III plays a role in proximal tubule dysfunction. Kidney Int 2008, 1999(74):52-61.
• [31]Pertovaara M, Bootorabi F, Kuuslahti M, Pasternack A, Parkkila S: Novel carbonic anhydrase autoantibodies and renal manifestations in patients with primary Sjogren’s syndrome. Rheumatology 2011, 50:1453-1457.
• [32]Nishita T, Matsushita H: Comparative immunochemical studies of carbonic anhydrase III in horses and other mammalian species. Comp Biochem Physiol 1988, 91:91-96.