title: Preparation and structural analysis of actinidain-processed atelocollagen of yellowfin tuna (Thunnus albacares).
abstract: Pepsin-hydrolyzed collagen (atelocollagen) is a trimer, consisting of alpha 1 and alpha 2 monomers, and shows molecular species corresponding to a monomer, dimer (beta chain), and trimer (gamma chain) by SDS-polyacrylamide gel electrophoresis. Atelocollagen was purified from yellowfin tuna (Thunnus albacares) by salt precipitation and cation-exchange chromatography. Enzymatic hydrolysis of the atelocollagen by actinidain, a cysteine protease purified from kiwifruit, was analyzed by SDS-polyacrylamide gel electrophoresis. The triple helical structure unique to collagen was retained in the atelocollagen as judged by circular dichroism spectra. The actinidain-processed atelocollagen showed only monomeric alpha 1 and alpha 2 chains, with no beta and gamma chains, by SDS-polyacrylamide gel electrophoresis; nevertheless, it retained the typical triple helical structure. It is suggested that actinidain cleaved the atelocollagen molecule at specific sites on the inside of the inter-strand cross-linking peptides.
authors.affiliation: Department of Biotechnological Science, Kinki University, Naga-gun, Wakayama, Japan. morimoto@bio.waka.kindai.ac.jp
authors.forename: Koichi
authors.initials: K
authors.lastname: Morimoto
authors.forename: Saori
authors.initials: S
authors.lastname: Kunii
authors.forename: Kaori
authors.initials: K
authors.lastname: Hamano
authors.forename: Ben'ichiro
authors.initials: B
authors.lastname: Tonomura
mesh_terms: D029042:Actinidia / Q000201:enzymology; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D002942:Circular Dichroism; D003094:Collagen / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005638:Fruit / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D013696:Temperature; D014413:Tuna
chemical_list: C059821:atelocollagen; D003094:Collagen; D003546:Cysteine Endopeptidases; C018745:actinidain