Identification of collagen types in tissues using HPLC-MS/MS.
A method for the determination and quantification of collagen types I-V in rat tissues has been developed. This method is based on collagen fragmentation by cyanogen bromide followed by trypsin digestion. After that, HPLC-MS/MS (HPLC coupled to an IT mass spectrometer) analyses of the resulting peptide mixtures (peptide maps) were performed. Specific peptides for each collagen type were selected. According to online databases, these peptides are present in human, bovine, and rat collagens. As a result, this method can be potentially applied to other species' tissues as well, such as human tissues, and provides a universal and simple method of quantifying collagen types. The applicability of this method for analyzing collagen types was demonstrated on rat tissues (skin, tendon, and aorta).
Pataridis
Statis
S
Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic. pataridis@biomed.cas.cz
Eckhardt
Adam
A
Mikulíková
Katerina
K
Sedláková
Pavla
P
Miksík
Ivan
I
D000595:Amino Acid Sequence; D000818:Animals; D002138:Calibration; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003094:Collagen / Q000032:analysis* / Q000235:genetics; D006801:Humans; D008126:Lod Score; D020033:Protein Isoforms / Q000032:analysis* / Q000235:genetics; D051381:Rats; D053719:Tandem Mass Spectrometry; D014020:Tissue Extracts / Q000737:chemistry*
D020033:Protein Isoforms; D014020:Tissue Extracts; D003094:Collagen