Langrock Tobias, García-Villar Natividad, Hoffmann Ralf
Institute of Bioanalytical Chemistry, Center for Biotechnology and Biomedicine (BBZ), Faculty of Chemistry and Mineralogy, University of Leipzig, Deutscher Platz 5, 04103 Leipzig, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Mar 1;847(2):282-8. doi: 10.1016/j.jchromb.2006.10.015. Epub 2006 Nov 7.
Collagens, the most abundant mammalian proteins, contain a high content of hydroxylated amino acids, such as, 3- and 4-cis-/trans-hydroxyproline (Hyp) and 5-hydroxylysine (Hyl). Whereas the global content of 4-Hyp was studied by amino acid analysis, no technique to determine all five hydroxyamino acids simultaneously in collagens has been reported. Here, we report the separation of all five hydroxyamino acids as well as two Hyp epimers from all other proteinogenic amino acids after derivatization with N(2)-(5-fluoro-2,4-dinitrophenyl)-l-valine amide (l-FDVA) by RPC-UV-ESI-MS. The general applicability of this method is shown for three Hyp-containing peptides as well as collagen type I.
胶原蛋白是哺乳动物中含量最丰富的蛋白质，含有高含量的羟基化氨基酸，如3-和4-顺式/反式-羟基脯氨酸（Hyp）以及5-羟基赖氨酸（Hyl）。虽然通过氨基酸分析研究了4-Hyp的总体含量，但尚未有同时测定胶原蛋白中所有五种羟基氨基酸的技术报道。在此，我们报告了在用N(2)-(5-氟-2,4-二硝基苯基)-L-缬氨酸酰胺（L-FDVA）衍生化后，通过反相色谱-紫外-电喷雾电离质谱法（RPC-UV-ESI-MS）从所有其他蛋白质ogenic氨基酸中分离出所有五种羟基氨基酸以及两种Hyp差向异构体。该方法对三种含Hyp的肽以及I型胶原蛋白的普遍适用性得到了证明。
