Cyanogen bromide peptides of the fibrillar collagens I, III, and V and their mass spectrometric characterization: detection of linear peptides, peptide glycosylation, and cross-linking peptides involved in formation of homo- and heterotypic fibrils.
The network of the fibrillar collagens I, III, and V, extracted from fetal calf skin and cleaved with cyanogen bromide, was studied by means of ultraviolet matrix-assisted laser desorption ionization time-of-flight mass spectrometry (UV-MALDI MS). Nearly all of the expected cyanogen bromide peptides of the different alpha chains were detected. Distinct peptides are identified that can serve as a reference signal for the individual alpha-chains. Homo- and heterotypic cross-linking patterns, some of which have not been described before for bovine collagen, are indicated by comparison of the mass spectrometric data with documented amino acid sequences. Potential cross-linking mechanisms are discussed. For example, the mass spectrometric data suggest that the formation of heterotypic I/III and I/V fibrils is substantially determined by the telo-regions of type I collagen, which are covalently connected to the corresponding helical and nonhelical cross-linking domains of adjacent molecules either by 4D or 0D-stagger bonds. The chemical nature of the cross-links can be concluded. The data also indicate a disturbed formation of heterotypic fibrils. Finally, collagen glycosylation can also be identified.
Henkel
Werner
W
Am Waldplatz 15, D-33098 Paderborn, Germany. Werner_Henkel@yahoo.de
Dreisewerd
Klaus
K
D000818:Animals; D002417:Cattle; D003094:Collagen / Q000737:chemistry; D024042:Collagen Type I / Q000737:chemistry*; D024061:Collagen Type III / Q000737:chemistry*; D024062:Collagen Type V / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003488:Cyanogen Bromide / Q000494:pharmacology*; D006031:Glycosylation; D013058:Mass Spectrometry / Q000379:methods; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014466:Ultraviolet Rays
D024042:Collagen Type I; D024061:Collagen Type III; D024062:Collagen Type V; D003432:Cross-Linking Reagents; D010455:Peptides; D011506:Proteins; D003094:Collagen; D003488:Cyanogen Bromide