title: The cleavage site preference of the porcine pepsin on the N-terminal α1 chain of bovine type I collagen: a focal analysis with mass spectrometry.
abstract: Bovine type I collagen consists of two α1 and one α2 chains, containing the internal triple helical regions and the N- and C-terminal telopeptides. In industries, it is frequently digested with porcine pepsin to produce a triple helical collagen without the telopeptides. However, the digestion mechanism is not precisely understood. Here, we performed a mass spectrometric analysis of the pepsin digest of the N-terminal telopeptide pQLSYGYDEKSTGISVP (1-16) in the α1 chain. When purified collagen was digested, pQLSYGY (1-6) and pQLSYGYDEKSTG (1-12) were identified, while DEKSTG (7-12) was not. When the N-terminal telopeptide mimetic synthetic peptide pQLSK(MOCAc)GYDEKSTGISK(Dnp)P-NH2was digested, pQLSK(MOCAc)GYDEKSTG (1-12) and ISK(Dnp)P-NH2(13-16) were readily identified, pQLSK(MOCAc)GY (1-6) and DEKSTGISK(Dnp)P-NH2(7-16) were weakly detected, and DEKSTG (7-12) was hardly identified. These results suggest that pepsin preferentially cleaves Tyr6-Asp7 and less preferentially Gly12-Ile13. They also suggest that the former cleavage requires native collagen structure, while the latter cleavage does not.
authors.affiliation: a Division of Food Science and Biotechnology , Graduate School of Agriculture, Kyoto University , Kyoto , Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Qian
authors.affiliation: b Medical Research Support Center, Graduate School of Medicine , Kyoto University , Kyoto , Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Ito
authors.affiliation: b Medical Research Support Center, Graduate School of Medicine , Kyoto University , Kyoto , Japan.
authors.forename: Junko
authors.initials: J
authors.lastname: Satoh
authors.affiliation: a Division of Food Science and Biotechnology , Graduate School of Agriculture, Kyoto University , Kyoto , Japan.
authors.forename: Hongmin
authors.initials: H
authors.lastname: Geng
authors.affiliation: c Nippi Research Institute of Biomatrix , Toride , Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Tanaka
authors.affiliation: c Nippi Research Institute of Biomatrix , Toride , Japan.
authors.forename: Shunji
authors.initials: S
authors.lastname: Hattori
authors.affiliation: a Division of Food Science and Biotechnology , Graduate School of Agriculture, Kyoto University , Kyoto , Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kojima
authors.affiliation: a Division of Food Science and Biotechnology , Graduate School of Agriculture, Kyoto University , Kyoto , Japan.
authors.forename: Teisuke
authors.initials: T
authors.lastname: Takita
authors.affiliation: a Division of Food Science and Biotechnology , Graduate School of Agriculture, Kyoto University , Kyoto , Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Yasukawa
mesh_terms: D000818:Animals; D002417:Cattle; D024042:Collagen Type I / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D010434:Pepsin A / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry; D012867:Skin / Q000737:chemistry; D034421:Sus scrofa; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D024042:Collagen Type I; D010446:Peptide Fragments; D010434:Pepsin A
keywords: collagen; mass spectrometry; pepsin; telopeptide