Canine prosomatostatin: isolation of a cDNA, regulation of gene expression, and characterization of post-translational processing intermediates.
Somatostatin is a tetradecapeptide (SS-14) initially isolated from the hypothalamus that is also found in D cells of the stomach and pancreas where it exerts an inhibitory action on a variety of gastrointestinal functions. Since many of concepts important to an understanding of gastrointestinal physiology are derived from experiments in the dog we examined somatostatin gene expression and post-translational processing in the canine fundus, antrum and pancreas. The canine somatostatin cDNA which is highly homologous to other known mammalian somatostatins was used to examine somatostatin expression in isolated canine fundic D-cells. Somatostatin expression induced by cholecystokinin (10(-8) M) was inhibited by the somatostatin analog, octreotide (10(-7) M). To examine somatostatin processing in the canine gut we noted that synthesis of SS-14 and somatostatin octacosapeptide (SS-28) involves endoproteolytic cleavage of prosomatostatin (proSS) at both paired and single basic amino-acid residues, respectively. Antisera capable of recognizing the amino-terminal residues of SS-28, SS-28(1-14) and SS-28(1-12) were characterized and identified concentrations of SS-28(1-12) but not SS-28(1-14) in the fundus, antrum and pancreas equivalent to those of SS-14. Since previous biosynthetic studies in canine fundic D-cells showed that SS-14 was synthesized without the appearance of a SS-28 intermediate, we hypothesize that proSS is sequentially cleaved at a dibasic site to produce SS-14 followed by monobasic cleavage that results in the formation of SS-28(1-12). Furthermore, equivalent amounts of SS-14 and SS-28(1-12) were co-released from canine fundic D-cells by CCK (10(-8) M) suggesting that the generation of these products occurs within the same regulated pathway of secretion.
Dickinson
C J
CJ
Department of Pediatrics, University of Michigan Medical Center, Ann Arbor, USA.
DelValle
J
J
Todisco
A
A
Gantz
I
I
Tong
L
L
Finniss
S
S
Yamada
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D001667:Binding, Competitive; D015152:Blotting, Northern; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs; D005753:Gastric Mucosa / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics*; D007118:Immunoassay; D008969:Molecular Sequence Data; D010179:Pancreas / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013004:Somatostatin / Q000737:chemistry* / Q000378:metabolism
D000906:Antibodies; D018076:DNA, Complementary; D010446:Peptide Fragments; D011498:Protein Precursors; D013004:Somatostatin; C028456:prosomatostatin