Cross-linking and fluorescence of pyrene-labeled collagen.
The pyrene-labeling of acid-soluble (type I) and acid-insoluble collagens from young and old rat tail tendon has been investigated. The pyrene excimer fluorescence is associated with stabilized pyrene labels bound to two adjacent aldehydes in monomeric young collagens. Polymeric young collagens, as well as monomeric and polymeric old collagens, tend to lose this specific arrangement. This is shown by salt and new chromatographic fractionation of monomeric and polymeric collagens. During denaturation, pyrene labels are released from saturated aldehydes in both alpha 1 and alpha 2 chains. This unstable pyrene-labeling is stabilized by NaBH4 reduction of the hydrazone bonds between aldehyde groups and pyrene-containing hydrazines. This stabilization reveals that alpha 1 contains more aldehyde groups than does alpha 2 in young collagen. Pepsin-solubilized, acid-insoluble collagens are partly cross-linked and, like acid-soluble collagens, exhibit the fluorescence of pyrene aggregates probably located at unidentified cross-links, different from unsaturated aldehyde-containing cross-links in acid-soluble collagens.
D000375:Aging; D000818:Animals; D003094:Collagen / Q000302:isolation & purification; D011487:Protein Conformation; D011721:Pyrenes; D051381:Rats; D013050:Spectrometry, Fluorescence; D013623:Tail; D013710:Tendons / Q000254:growth & development*; D014783:Viscosity
D011721:Pyrenes; D003094:Collagen; C030984:pyrene