Direct evidence for two distinct prosomatostatin converting enzymes. Detection using a rapid, sensitive, and specific assay for propeptide converting enzymes.
Many bioactive peptides are initially synthesized via larger precursors from which they are released by proteolytic cleavage at basic amino acids. Some precursors contain more than one final product peptide, multiple copies of a single peptide, or both. Different product peptides can be produced from a common precursor in different tissues. It is not currently known whether this cell-type specific production of bioactive peptides is mediated by different, specific propeptide converting enzymes (PCEs) or by a small number of similar PCEs. To resolve this issue for the conversion of prosomatostatin, the processing of prosomatostatin-I (aPSS-I) and prosomatostatin-II (aPSS-II) to either somatostatin-14 (SS-14) or somatostatin-28 (aSS-28), respectively, was examined in anglerfish islets. Two distinct forms of PSS PCE activity were detected using a rapid, sensitive, and specific assay. Examination of the specificity of these two enzyme activities showed that one proteolytic activity performs the aPSS-I to SS-14 conversion, while the other protease liberates aSS-28 from aPSS-II. The SS-14-generating PCE also cleaves aPSS-II to produce [Tyr7,Gly10]SS-14 (a tetra-decapeptide analog of SS-14) and converts proinsulin to insulin. The aSS-28-generating PCE does not process proinsulin. These results provide direct evidence that different, specific PCEs are required for liberation of SS-14 and aSS-28 from their precursors.
D000818:Animals; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005399:Fishes; D007515:Islets of Langerhans / Q000201:enzymology*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D011384:Proinsulin / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D013004:Somatostatin / Q000378:metabolism*; D013379:Substrate Specificity
D007527:Isoenzymes; D011498:Protein Precursors; D013004:Somatostatin; C028456:prosomatostatin; D011384:Proinsulin; D010450:Endopeptidases