Purification, characterization, and spasmogenic activity of neurotensin from the toad Bufo marinus

Fiona J. Warner; Elizabeth Burcher; Robert Carraway; J. Michael Conlon

doi:10.1016/S0196-9781(98)00063-1

Purification, characterization, and spasmogenic activity of neurotensin from the toad Bufo marinus

Fiona J. Warner, Elizabeth Burcher, Robert Carraway, J. Michael Conlon

School of Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

Neurotensin (NT) was isolated from an extract of the intestine of the cane toad, Bufo marinus and its primary structure established as: pGlu-Ala- Ile-Val-Ser-Lys-Ala-Arg-Arg-Pro-Tyr-Ile-Leu. This amino acid sequence shows five substitutions (Leu² → Ala, Tyr³ → Ile, Glu⁴ → Val, Asn⁵ → Ser, and Pro⁷ → Ala) compared with bovine NT. Synthetic Bufo NT (pD₂ = 8.05 ± 0.28) was equipotent and equally effective as bovine NT (pD₂ = 8.24 ± 0.38) in producing spasmogenic contraction of isolated segments of toad small intestine. However, the maximum response produced by Bufo NT was only 35 ± 2% of that produced by substance P. The potencies, but not the maximum responses, to Bufo and bovine NT were significantly (p < 0.05) attenuated by pre-treatment with atropine but neither parameter was significantly diminished by tetrodotoxin and indomethacin. The data suggest that the action of NT involves interaction with receptors on toad intestinal smooth muscle that recognize the C-terminal region of NT (residues 8-13) that has been fully conserved during evolution of tetrapods. Contractile activity is mediated, at least in part, by release of acetylcholine.

Original language	English
Pages (from-to)	1255-1261
Number of pages	7
Journal	Peptides
Volume	19
Issue number	7
DOIs	https://doi.org/10.1016/S0196-9781(98)00063-1
Publication status	Published (in print/issue) - Jul 1998

Keywords

Amphibian
Intestine
LANT-6
Myotropic
Neurotensin

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10.1016/S0196-9781(98)00063-1

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title = "Purification, characterization, and spasmogenic activity of neurotensin from the toad Bufo marinus",

abstract = "Neurotensin (NT) was isolated from an extract of the intestine of the cane toad, Bufo marinus and its primary structure established as: pGlu-Ala- Ile-Val-Ser-Lys-Ala-Arg-Arg-Pro-Tyr-Ile-Leu. This amino acid sequence shows five substitutions (Leu2 → Ala, Tyr3 → Ile, Glu4 → Val, Asn5 → Ser, and Pro7 → Ala) compared with bovine NT. Synthetic Bufo NT (pD2 = 8.05 ± 0.28) was equipotent and equally effective as bovine NT (pD2 = 8.24 ± 0.38) in producing spasmogenic contraction of isolated segments of toad small intestine. However, the maximum response produced by Bufo NT was only 35 ± 2% of that produced by substance P. The potencies, but not the maximum responses, to Bufo and bovine NT were significantly (p < 0.05) attenuated by pre-treatment with atropine but neither parameter was significantly diminished by tetrodotoxin and indomethacin. The data suggest that the action of NT involves interaction with receptors on toad intestinal smooth muscle that recognize the C-terminal region of NT (residues 8-13) that has been fully conserved during evolution of tetrapods. Contractile activity is mediated, at least in part, by release of acetylcholine.",

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AU - Conlon, J. Michael

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N2 - Neurotensin (NT) was isolated from an extract of the intestine of the cane toad, Bufo marinus and its primary structure established as: pGlu-Ala- Ile-Val-Ser-Lys-Ala-Arg-Arg-Pro-Tyr-Ile-Leu. This amino acid sequence shows five substitutions (Leu2 → Ala, Tyr3 → Ile, Glu4 → Val, Asn5 → Ser, and Pro7 → Ala) compared with bovine NT. Synthetic Bufo NT (pD2 = 8.05 ± 0.28) was equipotent and equally effective as bovine NT (pD2 = 8.24 ± 0.38) in producing spasmogenic contraction of isolated segments of toad small intestine. However, the maximum response produced by Bufo NT was only 35 ± 2% of that produced by substance P. The potencies, but not the maximum responses, to Bufo and bovine NT were significantly (p < 0.05) attenuated by pre-treatment with atropine but neither parameter was significantly diminished by tetrodotoxin and indomethacin. The data suggest that the action of NT involves interaction with receptors on toad intestinal smooth muscle that recognize the C-terminal region of NT (residues 8-13) that has been fully conserved during evolution of tetrapods. Contractile activity is mediated, at least in part, by release of acetylcholine.

AB - Neurotensin (NT) was isolated from an extract of the intestine of the cane toad, Bufo marinus and its primary structure established as: pGlu-Ala- Ile-Val-Ser-Lys-Ala-Arg-Arg-Pro-Tyr-Ile-Leu. This amino acid sequence shows five substitutions (Leu2 → Ala, Tyr3 → Ile, Glu4 → Val, Asn5 → Ser, and Pro7 → Ala) compared with bovine NT. Synthetic Bufo NT (pD2 = 8.05 ± 0.28) was equipotent and equally effective as bovine NT (pD2 = 8.24 ± 0.38) in producing spasmogenic contraction of isolated segments of toad small intestine. However, the maximum response produced by Bufo NT was only 35 ± 2% of that produced by substance P. The potencies, but not the maximum responses, to Bufo and bovine NT were significantly (p < 0.05) attenuated by pre-treatment with atropine but neither parameter was significantly diminished by tetrodotoxin and indomethacin. The data suggest that the action of NT involves interaction with receptors on toad intestinal smooth muscle that recognize the C-terminal region of NT (residues 8-13) that has been fully conserved during evolution of tetrapods. Contractile activity is mediated, at least in part, by release of acetylcholine.

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Purification, characterization, and spasmogenic activity of neurotensin from the toad Bufo marinus

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