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

Dongwuxue Yanjiu. 2015 May 18;36(3):174-7.

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 5.0 x 10⁻¹¹ mol/L and inducing maximal contraction at a concentration of 2.0 x 10⁻⁶ 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.

Keywords: Amphibian; Cholecystokinin; Salamander; Skin.

MeSH terms

  • Amino Acid Sequence
  • Amphibian Proteins / chemistry
  • Amphibian Proteins / genetics
  • Amphibian Proteins / isolation & purification*
  • Amphibian Proteins / pharmacology*
  • Animals
  • Base Sequence
  • Caudata / genetics*
  • Caudata / metabolism
  • Cholecystokinin / chemistry
  • Cholecystokinin / genetics
  • Cholecystokinin / isolation & purification*
  • Cholecystokinin / pharmacology*
  • Female
  • Male
  • Molecular Sequence Data
  • Muscle Contraction / drug effects
  • Muscle, Smooth / drug effects
  • Muscle, Smooth / physiology
  • Skin / chemistry*
  • Skin / metabolism
  • Swine

Substances

  • Amphibian Proteins
  • Cholecystokinin

Grants and funding

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)