Effect of oxazolidine E on collagen fibril formation and stabilization of the collagen matrix.
Oxazolidine E, an aldehydic cross-linking agent, is used to impart hydrothermal stability to collagen. The purpose of this study was to investigate the exact nature of oxazolidine E induced cross-links with collagen by using synthetic peptides having sequence homology with collagen type I. Tandem mass spectrometry revealed the formation of methylol and Schiff-base adducts upon reaction of oxazolidine E with the peptides. This was confirmed by allowing the reaction to proceed under reducing conditions using cyanoborohydride. Mass spectrometry (MS)-MS analysis clearly showed interaction of tryptophan and lysine residues with oxazolidine E and demonstrated that arginine could be cross-linked with glycine in the presence of oxazolidine E through the formation of a methylene bridge. Collagen fibrils regenerated from monomers in the presence and absence of oxazolidine E were studied using atomic force microscopy to investigate morphological alterations. Regenerated fibrils showing the typical 65 nm D-banding pattern were obtained from those formed both in the presence and absence of oxazolidine E, and there was no evidence of a change in the D-periodicity of these fibrils. This indicated that oxazolidine E does not hinder collagen molecules from correctly aligning to form the quarter-stagger structure.
Choudhury
Santanu Deb
SD
Institute of Molecular Biosciences, Massey University, Private Bag 11222, Palmerston North, New Zealand.
Haverkamp
Richard G
RG
DasGupta
Samir
S
Norris
Gillian E
GE
D000595:Amino Acid Sequence; D003094:Collagen / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004355:Drug Stability; D008855:Microscopy, Electron, Scanning; D010080:Oxazoles / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003432:Cross-Linking Reagents; D010080:Oxazoles; D010455:Peptides; C064210:oxazolidine; D003094:Collagen