Methylglyoxal inhibits the binding step of collagen phagocytosis.
Bacterial infection-induced fibrosis affects a wide variety of tissues, including the periodontium, but the mechanisms that dysregulate matrix turnover and mediate fibrosis are not defined. Since collagen turnover by phagocytosis is an important pathway for matrix remodeling, we studied the effect of the bacterial and eukaryotic cell metabolite, methylglyoxal (MGO), on the binding step of phagocytosis by periodontal fibroblasts. Type 1 collagen was treated with various concentrations of methylglyoxal, an important glucose metabolite that modifies Arg and Lys residues. The extent of MGO-induced modifications was authenticated by amino acid analysis, solubility, and cross-linking. Cells were incubated with fluorescent beads coated with collagen, and the percentage of phagocytic cells was estimated by flow cytometry. MGO inhibited collagen binding (20% of control for 10 mm MGO) in a time- and concentration-dependent manner. MGO-induced inhibition of binding was prevented by aminoguanidine, which blocks the formation of collagen cross-links. MGO reduced collagen binding strength and blocked intracellular calcium signaling. MGO modified the Arg residue in the critical alpha2beta1 integrin-binding recognition sequence of triple helical collagen peptides, whereas MGO-induced cross-linking of Lys residues played only a small role in binding inhibition. Thus, MGO modifications of Arg residues in collagen could be a key factor in the impaired degradation of collagen that promotes fibrosis in chronic infections, such as periodontitis.
Chong
Sandra A C
SA
Canadian Institutes of Health Research Group in Matrix Dynamics, University of Toronto, Toronto, Ontario M5S 3E2, Canada.
Lee
Wilson
W
Arora
Pam D
PD
Laschinger
Carol
C
Young
Edmond W K
EW
Simmons
Craig A
CA
Manolson
Morris
M
Sodek
Jaro
J
McCulloch
Christopher A
CA
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000737:chemistry; D002448:Cell Adhesion; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D038982:Integrin alpha2beta1 / Q000378:metabolism; D008239:Lysine / Q000737:chemistry; D010518:Periodontitis / Q000378:metabolism; D010587:Phagocytosis; D011485:Protein Binding; D011765:Pyruvaldehyde / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D015398:Signal Transduction
D000596:Amino Acids; D038982:Integrin alpha2beta1; D011765:Pyruvaldehyde; D003094:Collagen; D001120:Arginine; D008239:Lysine