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HORMONES AND SIGNALING

Identification of nonsulfated cholecystokinin-58 in canine intestinal extracts and its biological properties

¹CURE, Digestive Diseases Research Center, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles 90073; and Digestive Diseases Division, University of California Los Angeles School of Medicine, Los Angeles 90024; ³Section of Neurobiology, Physiology and Behavior, University of California, Davis 95616; ⁴Division of Immunology, Beckman Research Institute of the City of Hope, Duarte, California 91010; ²Division of Gastroenterology, Duke University Medical Center and Durham Veterans Affairs Medical Centers, Durham, North Carolina 22210; and ⁵University of Texas Health Science Center at San Antonio, Department of Physiology, San Antonio, Texas 78229

Submitted 15 December 2003 ; accepted in final form 8 February 2004

Nonsulfated CCK₅₈ [CCK₅₈(ns)] has not been considered to be of biological importance because CCK₅₈(ns) binds poorly to the CCK_A receptor and has only been identified once in intestinal extracts. In this work, a radioimmunoassay specific for the COOH-terminal region of gastrin and CCK (antibody 5135) was used to monitor the purification of CCK molecular forms from canine intestinal extracts. A minor immunoreactive peak was associated with a major absorbance peak during an ion-exchange, HPLC step. Characterization of this minor immunoreactive peak demonstrated that it was CCK₅₈(ns). CCK₅₈(ns) is 14% as immunoreactive as sulfated CCK₈ [CCK₈(s)]. Amino acid analysis demonstrated that CCK₅₈(ns) was present at 50% the amount of CCK₅₈(s). In addition, we found that CCK₅₈(ns) does not potently displace an ¹²⁵I-labeled CCK₁₀ analog from the CCK_A receptor in mouse pancreatic membranes and does not stimulate amylase release from isolated pancreatic acini, or stimulate pancreatic secretion in an anesthetized rat model. By contrast, CCK₅₈(ns) does bind to CCK_B receptors and stimulates gastric acid secretion via this receptor. The presence of CCK₅₈(ns) and its ability to selectively stimulate the CCK_B receptor without stimulation of the CCK_A receptor suggest that CCK₅₈(ns) may have unique physiological properties, especially tissues where the nonsulfated peptide can act as a paracrine or neurocrine agent.

gallbladder; pancreas; gastric acid; receptor binding; CCK_A; CCK_B

This article has been cited by other articles:

				S. V. Wu, K. G. Harikumar, R. J. Burgess, J. R. Reeve Jr., and L. J. Miller Effects of cholecystokinin-58 on type 1 cholecystokinin receptor function and regulation Am J Physiol Gastrointest Liver Physiol, September 1, 2008; 295(3): G641 - G647. [Abstract] [Full Text] [PDF]