Isolation and characterization of sulphated and nonsulphated forms of cholecystokinin-58 and their action on gallbladder contraction

Valentina Bonetto, Hans Jörnvall, Mats Andersson, Staffan Renlund, Viktor Mutt, Rannar Sillard

Research output: Contribution to journalArticle

Abstract

Cholecystokinin (CCK) exists in multiple molecular forms with different polypeptide lengths and the absence o-presence of sulphation. We have isolated sulphated and nonsulphated forms of CCK-58 from porcine intestine and have determined their bioactivities in a guinea-pig gallbladder contraction assay. Both forms co-eluted in cation-exchange chromatography and in several rounds of reverse-phase (RP)-HPLC, but separated upon RP-HPLC using a water/acetonitrile system with heptafluorobutyric acid as counter ion. Nonsulphated CCK-58 was the form detected by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry because of desulphation in that process. The biological activity of CCK-58 and CCK-33 is equipotent, although the kinetics of the response differ. Sulphated CCK-58 was found to be 35 times more potent than nonsulphated CCK-58. In contrast, sulphated CCK- 8 is 150 times more potent than nonsulphated CCK-8, and for sulphated and nonsulphated CCK-33, the activities differ by a factor of 100. This type of correlation indicates that the N-terminal end of CCK-58 partially compensates for the decrease in activity arising from the lack of sulphated tyrosine. Given its fairly high bioactivity, nonsulphated CCK-58 may have a physiological significance.

Original languageEnglish
Pages (from-to)336-340
Number of pages5
JournalEuropean Journal of Biochemistry
Volume264
Issue number2
DOIs
Publication statusPublished - Sep 1 1999

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Keywords

  • Cholecystokinin-58
  • Gallbladder contraction assay
  • MALDI mass spectrometry

ASJC Scopus subject areas

  • Biochemistry

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