Generation of glycosylated remnant epitopes from human collagen type II by gelatinase B.
Gelatinase B/matrix metalloproteinase-9 (MMP-9) is an inflammatory mediator and effector. Considerable amounts of gelatinase B are released by neutrophils in the synovial cavity of patients with rheumatoid arthritis, and gelatinase B-deficient mice are resistant against antibody-induced arthritis. Native human collagen type II is susceptible to cleavage by various collagenases (MMP-1, MMP-8, and MMP-13), which cleave at a single position in the triple helix. Although the triple-helical structure may persist after this single cleavage, we show that gelatinase B degrades the resulting fragments into small remnant peptides. These were identified by mass spectrometry and Edman degradation. Localization of 31 cleavage sites shows that the immunodominant epitopes remain intact after cleavage and may become available, processed as antigens and presented in MHC-II molecules. Furthermore, most post-translational modifications were identified on the fragments, including nine glycosylation sites. In particular, it is shown for the first time by structural analysis that in natural human collagen II, lysines in the main immunodominant epitope are modified by partial hydroxylation and partial glycosylation. Determination of T-cell reactivity against such fragments indicates that, besides the two known main immunodominant epitopes, other glyco-epitopes may be present in collagen II. This reinforces the role of glycopeptide antigens in autoimmunity.
Laboratory of Immunobiology, Rega Institute, University of Leuven, 3000 Leuven, Belgium. philippe.vandensteen@rega.kuleuven.ac.be
P E
PE
Van den Steen
P
P
Proost
D D
DD
Brand
A H
AH
Kang
J
J
Van Damme
G
G
Opdenakker
D017951:Antigen Presentation; D024043:Collagen Type II / Q000276:immunology / Q000378:metabolism*; D006020:Glycopeptides / Q000032:analysis / Q000276:immunology; D006801:Humans; D016056:Immunodominant Epitopes / Q000032:analysis*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis / Q000276:immunology*; D011499:Protein Processing, Post-Translational; D013601:T-Lymphocytes / Q000276:immunology
D024043:Collagen Type II; D006020:Glycopeptides; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D020780:Matrix Metalloproteinase 9