title: Peptides derived by processing of rat prosomatostatin near the amino-terminus: characterization, tissue distribution, and release.
abstract: Mammalian prosomatostatin (pro-S) undergoes extensive processing at the C-terminal segment where the somatostatin-like biological activities (S-14 and S-28) reside. The recent discovery of pro-S-(1-10) (antrin) as a prominent mature product in the stomach suggests that pro-S may also be processed at the N-terminus. In the present study we have developed an antibody directed against the N-terminal segment of pro-S-(1-10) capable of detecting peptides extended at the C-terminus of pro-S-(1-10) to characterize N-terminal processing of rat pro-S. Specifically, we have 1) examined the relative abundance of pro-S-(1-10)-like immunoactivity [pro-S-(1-10)] in different somatostatin tissues as an index of tissue-specific N-terminal processing, 2) compared the concentrations of pro-S N- and C-terminal immunoreactive peptides, 3) used HPLC and region-specific RIAs directed against both the N- and C-terminal segments of pro-S to identify and characterize novel N-terminal peptides, 4) studied the tissue distribution and release of the N-terminal peptides; and 5) characterized and quantified a 7-kDa molecule equivalent to pro-S without the C-terminal S-28 sequence. Acetic acid (1 M)-pepstatin extracts of hypothalamus, cerebral cortex, antrum, jejunal mucosa, and pancreas were fractionated by reverse phase and gel permeation HPLCs. Whole tissue extracts as well as the column effluent were monitored by region-specific RIAs using antibodies against pro-S-(1-10), S-28-(1-12), and S-14. Other than the pancreas, all S-producing tissues were rich in pro-S-(1-10) LI. Its concentration was 1- to 4-fold lower than those of S-14 LI and S-28-(1-12) LI. Tissue pro-S-(1-10) LI was heterogeneous, consisting of at least eight molecular forms with respective mol wt of 1,000 (1 kDa), 1,500 (1.5 kDa), 2,500 (2.5 kDa), 3,500 (3.5 kDa), 4,500 (4.5 kDa), 7,000 (7 kDa), 8,000 (8 kDa), and 10,000 (10 kDa). Based on the simultaneous presence or absence of C-terminal immunoreactivity, the 10-kDa form corresponded to pro-S, 8 kDa to pro-S-(1-76), and 7 kDa to pro-S without the S-28 sequence. The predominant N-terminal forms corresponded to 1 kDa [pro-S-(1-10)] and 7 kDa. The 1-, 1.5-, 2.5-, and 7-kDa forms were identified as secretion products in portal blood or in medium from cultured 1027 B2 islet somatostatin cells.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Fraser Laboratories, McGill University, Montreal, Quebec, Canada.
authors.forename: S N
authors.initials: SN
authors.lastname: Rabbani
authors.forename: Y C
authors.initials: YC
authors.lastname: Patel
mesh_terms: D000818:Animals; D002540:Cerebral Cortex / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D008297:Male; D008968:Molecular Conformation; D009994:Osmolar Concentration; D010455:Peptides / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D013004:Somatostatin / Q000378:metabolism*; D014018:Tissue Distribution
chemical_list: D010455:Peptides; D011498:Protein Precursors; D013004:Somatostatin; C028456:prosomatostatin