Vimentin Is the Specific Target in Skin Glycation

Abstract

Until now, the glycation reaction was considered to be a nonspecific reaction between reducing sugars and amino groups of random proteins. We were able to identify the intermediate filament vimentin as the major target for the AGE modification N ⑀ -(carboxymethyl)lysine (CML) in primary human fibroblasts. This glycation of vimentin is neither based on a slow turnover of this protein nor on an extremely high intracellular expression level, but remarkably it is based on structural properties of this protein. Glycation of vimentin was predominantly detected at lysine residues located at the linker regions using nanoLC-ESI-MS/MS. This modification results in a rigorous redistribution of vimentin into a perinuclear aggregate, which is accompanied by the loss of contractile capacity of human skin fibroblasts. CMLinduced rearrangement of vimentin was identified as an aggresome. This is the first evidence that CML-vimentin represents a damaged protein inside the aggresome, linking the glycation reaction directly to aggresome formation. Strikingly, we were able to prove that the accumulation of modified vimentin can be found in skin fibroblasts of elderly donors in vivo, bringing AGE modifications in human tissues such as skin into strong relationship with loss of organ contractile functions.

Dicarbonyl-induced accelerated aging in vitro in human skin fibroblasts

“…In addition, GO has been shown to inhibit glutathione reductase (GR) (Shangari et al 2006), reducing the level of GSH and in turn GO and MGO detoxification by the cytosolic glyoxalase system. CML-modified proteins as well as other AGEs are good molecular markers of aging because they accumulate during normal aging in vivo (Hipkiss 2006;Kueper et al 2007). In our studies the observation that there is a significant increase in CML-proteins is additional evidence supporting the induction of accelerated aging in GO-and MGOtreated human fibroblasts.…”

Muscle Creatine Kinase Deficiency Triggers Both Actin Depolymerization and Desmin Disorganization by Advanced Glycation End Products in Dilated Cardiomyopathy

“…Partial colocalization of AGE adducts and desmin aggregates was reported in skeletal muscle fibers from patients suffering from desminopathies, suggesting that desmin is prone to AGE adduct modifications in the context of muscle disease (36). Interestingly, vimentin, another type III intermediate filament protein closely related to desmin, is also a preferential target of AGE in the senescent fibroblast where it participates in aggresome formation (37,38). Desmin and vimentin play an important role in mitochondria distribution in muscle cells and through their interaction with tubulin regulate mitochondrial affinity for ADP, V max of oxygen consumption, and creatine kinase coupling with the adenine nucleotide translocator (39 -41).…”

Effects of Glyceryl Glucoside on AQP3 Expression, Barrier Function and Hydration of Human Skin

“…Cells were stained as described previously [16]. Briefly, keratinocytes were fixed in 3% paraformaldehyde for 30 min at room temperature and permeabilized with 0.5% Triton X-100 (Sigma-Aldrich, St. Louis, Mo., USA) for 5 min.…”

“…Flow cytometry was performed according to a previously described protocol [16]. Briefly, cells were centrifuged at 13,000 rpm and the pellet was washed twice with PBS followed by incubation in 3% paraformaldehyde (Sigma-Aldrich).…”