title: Stable isotope-labeled collagen: a novel and versatile tool for quantitative collagen analyses using mass spectrometry.
abstract: Collagens are the most abundant proteins in animals and are involved in many physiological/pathological events. Although various methods have been used to quantify collagen and its post-translational modifications (PTMs) over the years, it is still difficult to accurately quantify type-specific collagen and minor collagen PTMs. We report a novel quantitative method targeting collagen using stable isotope-labeled collagen named "SI-collagen", which was labeled with isotopically heavy lysine, arginine, and proline in fibroblasts culture. We prepared highly labeled and purified SI-collagen for use as an internal standard in mass spectrometric analysis, particularly for a new approach using amino acid hydrolysis. Our method enabled accurate collagen analyses, including quantification of (1) type-specific collagen (types I and III in this paper), (2) total collagen, and (3) collagen PTMs by LC-MS with high sensitivity. SI-collagen is also applicable to other diverse analyses of collagen and can be a powerful tool for various studies, such as detailed investigation of collagen-related disorders.
authors: Nippi Research Institute of Biomatrix , 520-11 Kuwabara, Toride, Ibaraki 302-0017, Japan.
authors: Yuki
authors: Y
authors: Taga
authors: Masashi
authors: M
authors: Kusubata
authors: Kiyoko
authors: K
authors: Ogawa-Goto
authors: Shunji
authors: S
authors: Hattori
mesh_terms: D000818:Animals; D001120:Arginine; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods; D003094:Collagen / Q000032:analysis*; D005347:Fibroblasts / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D008239:Lysine; D013058:Mass Spectrometry / Q000379:methods; D011392:Proline; D051381:Rats
chemical_list: D003094:Collagen; D001120:Arginine; D011392:Proline; D008239:Lysine