Conformational Dependence of Collagenase (Matrix Metalloproteinase-1) Up-regulation by Elastin Peptides in Cultured Fibroblasts

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Background:The interaction of the peptide VGVAPG with the elastin binding protein is critically involved in aneurysm progression.Results: A molecular model of this interaction is proposed and explored using a site-directed mutagenesis strategy. Conclusion: Three residues, Leu-103, Arg-107, and Glu-137, of elastin binding protein are critical players in this interaction. Significance: Our data now allow the design of antagonists of VGVAPG.

Section: Discussionmentioning

confidence: 67%

“…EBP is known to interact with various agonists (13,(31)(32)(33)(34)(35). In previous work, we have established that hexapeptides interacting with EBP share the XGXXPG consensus sequence that is thought to stabilize a type VIII ␤-turn (30,31).…”

Section: Discussionmentioning

confidence: 99%

Interaction between the Elastin Peptide VGVAPG and Human Elastin Binding Protein

Blanchevoye

Floquet

Scandolera

et al. 2013

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“…This was achieved using 1 M KOH in 80% aqueous ethanol (36). The obtained mixture of elastin peptides is termed -elastin (kE) and exhibits the same biological and physical properties as elastin hydrolysates (5).…”

Section: Methodsmentioning

confidence: 99%

“…However, when Senior et al (4) demonstrated that elastin digestion products were chemotactic for neutrophils and macrophages, it became suddenly apparent that peptides derived from amorphous elastin could modulate cell physiology. In fact, it has been shown because that fibroblasts (5)(6)(7)(8)(9)(10)(11)(12), smooth muscle cells (7,(13)(14)(15), endothelial cells (16 -19), leukocytes (20,21), and lymphocytes (22) were sensitive to the presence of these peptides yielding a broad range of biological activities (see Ref. 23 for a review).…”

mentioning

confidence: 99%

The Elastin Receptor Complex Transduces Signals through the Catalytic Activity of Its Neu-1 Subunit

Duca

Blanchevoye

Cantarelli

et al. 2007

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113

118

The binding of elastin peptides on the elastin receptor complex leads to the formation of intracellular signals but how this is achieved remains totally unknown. Using pharmacological inhibitors of the enzymatic activities of its subunits, we show here that the elastin peptide-driven ERK1/2 activation and subsequent pro-MMP-1 production, observed in skin fibroblasts when they are cultured in the presence of these peptides, rely on a membrane-bound sialidase activity. As lactose blocked this effect, the elastin receptor sialidase subunit, Neu-1, seemed to be involved. The use of a catalytically inactive form of Neu-1 and the small interfering RNA-mediated decrease of Neu-1 expression strongly support this view. Finally, we report that N-acetyl neuraminic acid can reproduce the effects of elastin peptides on both ERK1/2 activation and pro-MMP-1 production. Altogether, our results indicate that the enzymatic activity of the Neu-1 subunit of the elastin receptor complex is responsible for its signal transduction, presumably through sialic acid generation from undetermined substrates.Elastin is the extracellular matrix protein responsible for the elasticity of tissues. It is more abundant in tissues where resilience is required, such as skin, lung, ligaments, or large arteries (1). Elastin is constituted of tropoelastin molecules covalently bound to each other by covalent cross-links (2) and its hydrophobic and highly cross-linked nature make of it a very durable polymer experiencing essentially no turnover in healthy tissues (3).The biological role of elastin was originally thought to be restricted to this mechanical function. However, when Senior et al. (4) demonstrated that elastin digestion products were chemotactic for neutrophils and macrophages, it became suddenly apparent that peptides derived from amorphous elastin could modulate cell physiology. In fact, it has been shown because that fibroblasts (5-12), smooth muscle cells (7, 13-15), endothelial cells (16 -19), leukocytes (20, 21), and lymphocytes (22) were sensitive to the presence of these peptides yielding a broad range of biological activities (see Ref. 23 for a review). A corollary of these observations was that those cells do express a receptor for elastin peptides.The elastin receptor complex is constituted of three subunits, one peripheral 67-kDa subunit, which actually binds elastin, and two membrane-associated proteins of 61 and 55 kDa, respectively (10). The 67-kDa elastin-binding protein (EBP) 2 binds the VGVAPG elastin sequence with high affinity. Additionally, EBP can be eluted from elastin affinity column by galactosugars suggesting that the elastin-EBP interaction could be regulated by galactosugars bound on a lectin site on EBP (10, 24). Consequently, galactosugars such as lactose are commonly used antagonists of EBP.Later, the nature of this subunit was revealed by the work of Privitera et al. (25) who have shown that EBP is an enzymatically spliced variant of lysosomal ␤-galactosidase (␤-Gal, EC 3.2.1.23). Consequently, it was hypot...

“…Brassart and colleagues (32) reported that pertussis toxinsensitive G proteins and tyrosine kinase are involved in the EBP-mediated up-regulation of matrix metalloproteinase-2 in HT-1080 cancer cells. Because the abovementioned data clearly suggest that the cell surface EBP functions as a subunit of a true receptor capable of transmitting physiological signals from the extracellular matrix into the cell interior, the present studies have been aimed at a detailed disclosure of the intracellular signaling pathways generated by the association between elastin-derived peptides and the EBP.…”

mentioning

confidence: 99%

Signaling Pathways Transduced through the Elastin Receptor Facilitate Proliferation of Arterial Smooth Muscle Cells

Mochizuki

Brassart

Hinek

2002

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216

148

In this report we demonstrate that soluble peptides, elastin degradation products stimulate proliferation of arterial smooth muscle cells. We show that these effects are due to generation of intracellular signals transduced through the cell surface elastin receptor, which consists of peripheral 67-kDa elastin-binding protein (EBP) (spliced variant of ␤-galactosidase), immobilized to the transmembrane sialidase and the protective protein. We found that elastin receptor-transduced signaling triggers activation of G proteins, opening of L-type calcium channels, and a sequential activation of tyrosine kinases: FAK, c-Src, platelet-derived growth factorreceptor kinase and then Ras-Raf-MEK1/2-ERK1/2 phosphorylation cascade. This, in turn, causes an increase in expression of cyclins and cyclin-dependent kinases, and a consequent increase in cellular proliferation. The EBP-transduced signals also induce tyrosine kinase-dependent phosphorylation of ␤-tubulin, LC3, microtubule-associated protein 1, and ␣-actin and troponin-T, which could be linked to reorganization of cytoskeleton. We have also disclosed that induction of these signals can be abolished by anti-EBP antibody or by galactosugars, which cause shedding of EBP from the cell surface. Moreover, elastin-derived peptides did not induce proliferation of EBP-deficient cells derived from patients bearing a nonsense mutation of the ␤-galactosidase gene or sialidase-deficient cells from patients with congenital sialidosis.It has been well established that formation of neointima in vascular diseases is associated with impaired assembly of tropoelastin into insoluble elastin (1-6) and with extensive degradation of the elastin-rich extracellular matrix by numerous proteinases leaking from the serum and secreted from the infiltrating platelets, leukocytes, and activated vascular cells (7-10). It has also been suggested that local accumulation of non-assembled tropoelastin and small elastin-derived peptides may constitute an important factor in the activation of the normally quiescent medial SMC 1 into the proliferative and migratory phenotype, which participates in the formation of the occlusive neointima in vascular diseases (2,3,(11)(12)(13)(14)(15) Elastin does not contain the RGD sequence and does not interact with cell surface integrins. Our previous studies demonstrated that numerous cell types, including vascular myocytes, express the cell surface elastin receptor complex, which consists of three subunits (25,26), and that the average cell contains ϳ2 ϫ 10 6 elastin binding sites with the binding affinity (K d ) of 8 nM (27). We found that two of those subunits (55-and 61-kDa) are anchored to the plasma membrane, whereas the third, a peripheral 67-kDa protein, actually binds elastin (25). This major functional component of the receptor complex was named the elastin binding protein (EBP). The repeat hexapeptide in tropoelastin, VGVAPG, has been identified as a chief ligand for high affinity binding to this cell surface receptor (25-27). It has been later established t...