5:15 PM - 5:30 PM
[2Ap-09] Biochemical Characterization of Major Collagen and its Constituent Subunits from Moon Jellyfish
Keywords:Collagen, Unused Marine Resources, Protein
Purpose:Jellyfish collagen is currently used in cosmetics and research reagents. Our group has developed a method to solubilize collagen from jellyfish tissue using endogenous proteases. The resulting endogenous protease-solubilized collagen (ESC) exhibits a distinct pattern on SDS-PAGE compared to vertebrate Type I collagen. However, little is known about the specific collagen molecular species within ESC and the properties of each constituent subunit. In this study, our aim was to extract ESC from moon jellyfish, isolate the major molecular species of collagen and elucidate its subunit composition.
Materials and Methods:Fresh moon jellyfish were frozen and subjected to autodigestion at low temperatures to obtain ESC. ESC or its thermally denatured product underwent cation exchange chromatography or gel filtration to isolate the major molecular species or constituent subunits of collagen. The resulting collagen or subunits were then analyzed using SDS-PAGE, amino acid analysis, or peptide mapping for characterization.
Results:On SDS-PAGE, ESC showed distinct bands at 150-160 K, 80-90 K, 50 K, and 25 K, indicating a collagen-like amino acid composition. Cation exchange chromatography revealed that the major molecular species of collagen comprising ESC contained all the aforesaid components. Upon denaturation, ESC was fractionated into fractions A (25 K), B① (50 K), B② (80 K), C (150 K), D (80-90 K), and E (160 K). Amino acid analysis suggested that fractions A and B① contained low glycine content, indicating the presence of non-collagenous regions. These findings indicate successful isolation of the major collagen molecular species in ESC and suggest the possibility of non-collagenous regions in some constituent subunits.
Materials and Methods:Fresh moon jellyfish were frozen and subjected to autodigestion at low temperatures to obtain ESC. ESC or its thermally denatured product underwent cation exchange chromatography or gel filtration to isolate the major molecular species or constituent subunits of collagen. The resulting collagen or subunits were then analyzed using SDS-PAGE, amino acid analysis, or peptide mapping for characterization.
Results:On SDS-PAGE, ESC showed distinct bands at 150-160 K, 80-90 K, 50 K, and 25 K, indicating a collagen-like amino acid composition. Cation exchange chromatography revealed that the major molecular species of collagen comprising ESC contained all the aforesaid components. Upon denaturation, ESC was fractionated into fractions A (25 K), B① (50 K), B② (80 K), C (150 K), D (80-90 K), and E (160 K). Amino acid analysis suggested that fractions A and B① contained low glycine content, indicating the presence of non-collagenous regions. These findings indicate successful isolation of the major collagen molecular species in ESC and suggest the possibility of non-collagenous regions in some constituent subunits.