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Am J Physiol Gastrointest Liver Physiol 276: G808-G816, 1999;
0193-1857/99 $5.00
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Vol. 276, Issue 4, G808-G816, April 1999

COOH-terminally extended secretins are potent stimulants of pancreatic secretion

Travis E. Solomon1, John H. Walsh1, Louis Bussjaeger2, Yumei Zong1, James W. Hamilton2, F. J. Ho1, Terry D. Lee3, and Joseph R. Reeve Jr.1

1 CURE: Digestive Diseases Research Center, Greater Los Angeles Department of Veterans Affairs Health Care System, Los Angeles 90073; Digestive Diseases Division, University of California, Los Angeles, School of Medicine, Los Angeles, California 90024; 2 Kansas City Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128; Department of Biochemistry, Kansas University Medical Center, Kansas City, Kansas 66160; and 3 Division of Immunology, Beckman Research Institute, The City of Hope Research Institute, Duarte, California 91010

Posttranslational processing of preprosecretin generates several COOH-terminally extended forms of secretin and alpha -carboxyl amidated secretin. We used synthetic canine secretin analogs with COOH-terminal -amide, -Gly, or -Gly-Lys-Arg to examine the effects of COOH-terminal extensions of secretin on bioactivity and detection in RIA. Synthetic products were purified by reverse-phase and ion-exchange HPLC and characterized by reverse-phase isocratic HPLC and amino acid, sequence, and mass spectral analyses. Secretin and secretin-Gly were noted to coelute during reverse-phase HPLC. In RIA using eight different antisera raised against amidated secretin, COOH-terminally extended secretins had little or no cross-reactivity. Bioactivity was assessed by measuring pancreatic responses in anesthetized rats. Amidated canine and porcine secretins were equipotent. Secretin-Gly and secretin-Gly-Lys-Arg had potencies of 81 ± 9% (P > 0.05) and 176 ± 13% (P < 0.01), respectively, compared with amidated secretin, and the response to secretin-Gly-Lys-Arg lasted significantly longer. These data demonstrate that 1) amidated secretin and secretin-Gly are not separable under some chromatographic conditions, 2) current RIA may not detect bioactive COOH-terminally extended forms of secretin in tissue extracts or blood, and 3) the secretin receptor mediating stimulation of pancreatic secretion recognizes both amidated and COOH-terminally extended secretins.

posttranslational processing; amidation; radioimmunoassay; physiology


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