Purification and Characterization of the Main Pepsinogen from the Shark, Centroscymnus coelolepis

• Published in: Journal of biochemistry (Tokyo) Vol. 124; no. 2; pp. 287 - 293
• Main Authors: Nguyen, Anh-Dung, Nungaray, Jesus, Martel, Annie, Le Goffic, Francois, Mollé, Daniel, Léonil, Joelle
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.08.1998
• Subjects: Amino Acid Sequence; Animals; Autolysis; Catalysis; characterization; Endopeptidases - metabolism; Gastric Mucosa - enzymology; Insulin - metabolism; Molecular Sequence Data; pepsin; Pepsinogen A - chemistry; Pepsinogen A - isolation & purification; Pepsinogen A - metabolism; Sequence Analysis; shark; Sharks; specificity; Substrate Specificity
• ISSN: 0021-924X
• DOI: 10.1093/oxfordjournals.jbchem.a022109

The main pepsinogen from the mucosa of the shark, Centroscymnus coelolepis, has been purified and characterized. This zymogen, the most abundant protein in terms of quantity and activity (yield 72%), is a homogeneous monomer of molecular weight 42±0.7 kDa, as determined by electrophoresis. The aspartyl proteinase nature of this enzyme was confirmed by the considerable inhibition by pepstatin. Its specificity as to the oxidized B-chain of bovine insulin was determined using electrospray ionization mass spectrometry (ESI-MS) coupled with reversed phase high pressure liquid chromatography (RP-HPLC). The 15–16 Leu-Tyr bond was rapidly cleaved in this substrate, followed by the 24–25 Phe-Phe, 25–26 Phe-Tyr, and 11–12 Leu-Val bonds.