Purification and characterization of cholecystokinin from the skin of salamander <em>Tylototriton verrucosus</em>

Wen-Bin JIANG; Ma HAKIM; Lei LUO; Bo-Wen LI; Shi-Long YANG; Yu-Zhu SONG; Ren LAI; Qiu-Min LU

Volume 36 Issue 3

May 2015

Turn off MathJax

Article Contents

Article Navigation > Zoological Research > 2015 > 36(3): 174-177

Wen-Bin JIANG, Ma HAKIM, Lei LUO, Bo-Wen LI, Shi-Long YANG, Yu-Zhu SONG, Ren LAI, Qiu-Min LU. Purification and characterization of cholecystokinin from the skin of salamander Tylototriton verrucosus. Zoological Research, 2015, 36(3): 174-177.

Citation:

Wen-Bin JIANG, Ma HAKIM, Lei LUO, Bo-Wen LI, Shi-Long YANG, Yu-Zhu SONG, Ren LAI, Qiu-Min LU. Purification and characterization of cholecystokinin from the skin of salamander Tylototriton verrucosus. Zoological Research, 2015, 36(3): 174-177.

Citation:

PDF( 355 KB)

Purification and characterization of cholecystokinin from the skin of salamander Tylototriton verrucosus

1 Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming Yunnan 650500, China;
2 Key Laboratory of Animal Models and Human Disease Mechanisms of Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Kunming Yunnan 650223, China

Funds: This work was supported by the National Basic Research Program of China (973 Program) (2013CB911300), National Natural Science Foundation of China (U1132601) and the Chinese Academy of Sciences (SAJC201308)

More Information

Corresponding author: Ren LAI, Qiu-Min LU
Received Date: 2015-03-16
Rev Recd Date: 2015-04-09
Publish Date: 2015-05-08

Abstract

Abstract

As a group of intestinal hormones and neurotransmitters, cholecystokinins (CCKs) regulate and affect pancreatic enzyme secretion, gastrointestinal motility, pain hypersensitivity, digestion and satiety, and generally contain a DYMGWMDFG sequence at the C-terminus. Many CCKs have been reported in mammals. However, only a few have been reported in amphibians, such as Hyla nigrovittata, Xenopus laevis, and Rana catesbeiana, with none reported in urodele amphibians like newts and salamanders. Here, a CCK called CCK-TV was identified and characterized from the skin of the salamander Tylototriton verrucosus. This CCK contained an amino acid sequence of DYMGWMDF-NH₂ as seen in other CCKs. A cDNA encoding the CCK precursor containing 129 amino acid residues was cloned from the cDNA library of T. verrucosus skin. The CCK-TV had the potential to induce the contraction of smooth muscle strips isolated from porcine gallbladder, eliciting contraction at a concentration of 5.0x10^-11 mol/L and inducing maximal contraction at a concentration of 2.0x10^-6 mol/L. The EC₅₀ was 13.6 nmol/L. To the best of our knowledge, this is the first report to identify the presence of a CCK in an urodele amphibian.
- Cholecystokinin,
- Salamander,
- Skin,
- Amphibian

FullText(HTML)

References(19)

References

[1]	Bevins CL, Zasloff M. 1990. Peptides from frog skin. Annual Review of Biochemistry, 59: 395-414.
[2]	Dimaline R, Lee CM. 1990. Chicken gastrin: A member of the gastrin/CCK family with novel structure-activity relationships. The American Journal of Physiology, 259(5 Pt 1): G882-G888.
[3]	Erspamer V, Melchiorri P, Broccardo M, Erspamer GF, Falaschi P, Improta G, Negri L, Renda T. 1981. The brain-gut-skin triangle: new peptides. Peptides, 2(Suppl. 2): 7-16.
[4]	Johnsen AH, Rehfeld JF. 1992. Identification of cholecystokinin/gastrin peptides in frog and turtle. Evidence that cholecystokinin is phylogenetically older than gastrin. European Journal of Biochemistry, 207(2): 419-428.
[5]	Johnsen AH. 1994. Identification of cholecystokinin from frog and turtle. Divergence of cholecystokinin and gastrin occurred before the evolution of amphibia. European Journal of Biochemistry, 224(2): 691-702.
[6]	Lai R, Liu H, Lee W H, Zhang Y. 2001. A novel bradykinin related peptide from skin secretions of toad Bombina maxima and its precursor containing six identical copies of the final product. Biochemical and Biophysical Research Communications, 286(2): 259-263.
[7]	Lai R, Liu H, Lee WH, Zhang Y. 2002. A novel proline rich bombesin-related peptide (PR-bombesin) from toad Bombina maxima. Peptides, 23(3): 437-442.
[8]	Liu X H, Wang Y P, Cheng L H, Song Y Z, Lai R. 2007. Isolation and cDNA cloning of cholecystokinin from the skin of Rana nigrovittata. Peptides, 28(8): 1540-1544.
[9]	Mu L, Tang J, Liu H, Shen C, Rong M, Zhang Z, Lai R. 2014. A potential wound-healing-promoting peptide from salamander skin. The FASEB Journal, 2014, 28(9): 3919-3929.
[10]	Lu QM, Lai R, Zhang Y. 2010. Animal toxins and human disease: from single component to venomics, from biochemical characterization to disease Mechanisms, from crude venom utilization to rational drug design. Zoological Research, 31(1): 2-16. (in Chinese)
[11]	Nielsen KG, Bomgren P, Holmgren S, Johnsen AH. 1998. Gastrin and cholecystokinin of the bullfrog, Rana catesbeiana, have distinct effects on gallbladder motility and gastric acid secretion in vitro. General and Comparative Endocrinology, 112(2): 247-254.
[12]	Oliver AS, Vigna SR. 1996. CCK-X receptors in the endothermic mako shark (Isurus oxyrinchus). General and Comparative Endocrinology, 102(1): 61-73.
[13]	Oliver AS, Vigna SR. 1997. CCK-A- and CCK-B-like receptors in the gallbladder and stomach of the alligator (Alligator mississippiensis). General and Comparative Endocrinology, 105(1): 91-101.
[14]	Rourke IJ, Rehfeld JF, Møller M, Johnsen AH. 1997. Characterization of the cholecystokinin and gastrin genes from the bullfrog, Rana catesbeiana: evolutionary conservation of primary and secondary sites of gene expression. Endocrinology, 138(4): 1719-1727.
[15]	Wakabayashi T, Kato H, Tachibana S. 1985. Complete nucleotide sequence of mRNA for caerulein precursor from Xenopus skin: the mRNA contains an unusual repetitive structure. Nucleic Acids Research, 13(6): 1817-1828.
[16]	Wu J, Liu H, Yang H L, Yu H N, You D W, Ma Y F, Ye H H, Lai R. 2011. Proteomic analysis of skin defensive factors of tree frog Hyla simplex. Journal of Proteome Research, 10(9): 4230-4240.
[17]	Xu X Q, Lai R. 2015. The chemistry and biological activities of peptides from amphibian skin secretions. Chemical Reviews, 115(4): 1760-1846.
[18]	You D W, Hong J, Rong M Q, Yu H N, Liang S P, Ma Y F, Yang H L, Wu J, Lin D H, Lai R. 2009. The first gene-encoded amphibian neurotoxin. The Journal of Biological Chemistry, 284(33): 22079-22086.
[19]	Zhang Y. 2006. Amphibian skin secretions and bio-adaptive significance — Implications from Bombina maxima skin secretion proteome. Zoological Research, 27(1): 101-112. (in Chinese)