Proteolysis Regulates Exposure of the IIICS-1 Adhesive Sequence in Plasma Fibronectin

Ugarova, Tatiana P.; Ljubimov, Alexander V.; Deng, Lynn; Plow, Edward F.

doi:10.1021/bi960717s

Cited by 36 publications

(31 citation statements)

References 59 publications

(74 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…22,[24][25][26] Fibronectin proteolysis by chymotrypsin and cathepsins B and D exposes integrin-binding sites such as the RGD (Arg-GlyAsp) site thereby enhancing integrin binding. 24,56 It is currently unknown where exactly GzmB cleaves fibronectin, however, GzmB preferentially cleaves after Asp residues 57,58 and Buzza et al have demonstrated that GzmB cleaves vitronectin at the RGD integrin-binding site. 27 Although fibronectin proteolysis by some proteases can enhance integrin binding and contraction, 24,56 GzmB-mediated proteolysis may have profoundly different effects depending on the site of cleavage and, although not confirmed, could negatively influence contraction should integrin binding to the RGD site be disrupted.…”

Section: Discussionmentioning

confidence: 99%

“…24,56 It is currently unknown where exactly GzmB cleaves fibronectin, however, GzmB preferentially cleaves after Asp residues 57,58 and Buzza et al have demonstrated that GzmB cleaves vitronectin at the RGD integrin-binding site. 27 Although fibronectin proteolysis by some proteases can enhance integrin binding and contraction, 24,56 GzmB-mediated proteolysis may have profoundly different effects depending on the site of cleavage and, although not confirmed, could negatively influence contraction should integrin binding to the RGD site be disrupted. In our study, we demonstrate that HFD-fed AKO mice express GzmB within the granulation tissue, contain elevated levels of a fibronectin fragment similar to that generated by GzmB in vitro, which corresponds with significantly delayed wound contraction that was improved in DKO mice.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Granzyme B degrades extracellular matrix and contributes to delayed wound closure in apolipoprotein E knockout mice

Hiebert

Granville

2013

Cell Death Differ

View full text Add to dashboard Cite

Chronic inflammation and excessive protease activity have a major role in the persistence of non-healing wounds. Granzyme B (GzmB) is a serine protease expressed during chronic inflammation that, in conjunction with perforin, has a well-established role in initiating apoptotic cell death. GzmB is also capable of acting extracellularly, independent of perforin and can degrade several extracellular matrix (ECM) proteins that are critical during wound healing. We used apolipoprotein E (ApoE) knockout (AKO) mice as a novel model of chronic inflammation and impaired wound healing to investigate the role of GzmB in chronic wounds. Wild-type and AKO mice were grown to 7 weeks (young) or 37 weeks (old) of age on a regular chow or high-fat diet (HFD), given a 1-cm diameter full thickness wound on their mid dorsum and allowed to heal for 16 days. Old AKO mice fed a HFD exhibited reduced wound closure, delayed contraction, chronic inflammation and altered ECM remodeling. Conversely, GzmB/ApoE double knockout mice displayed improved wound closure and contraction rates. In addition, murine GzmB was found to degrade both fibronectin and vitronectin derived from healthy mouse granulation tissue. In addition, GzmB-mediated degradation of fibronectin generated a fragment similar in size to that observed in non-healing mouse wounds. These results provide the first direct evidence that GzmB contributes to chronic wound healing in part through degradation of ECM.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Granzyme B degrades extracellular matrix and contributes to delayed wound closure in apolipoprotein E knockout mice

Hiebert

Granville

2013

Cell Death Differ

View full text Add to dashboard Cite

show abstract

“…This integrin binds to the connecting segment region called IIICS or V (variable) as well as to a similar site in the Hep II domain of fibronectin (42,43). In native plasma fibronectin the CS-1 adhesion site is masked but can be exposed under partial proteolysis with cathepsin D (44). Because the ADAM13 adhesive domain can bind to the second heparin-binding domain of fibronectin and the ADAM13 metalloprotease domain can cleave fibronectin, it will be of interest to investigate whether this cleavage can also expose the CS-1 site.…”

Section: Discussionmentioning

confidence: 99%

ADAM13 Disintegrin and Cysteine-rich Domains Bind to the Second Heparin-binding Domain of Fibronectin

Gaultier¹,

Cousin²,

Darribère³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

ADAM13 is a member of the disintegrin and metalloprotease protein family that is expressed on cranial neural crest cells surface and is essential for their migration. ADAM13 is an active protease that can cleave fibronectin in vitro and remodel a fibronectin substrate in vivo. Using a recombinant secreted protein containing both disintegrin and cysteine-rich domains of ADAM13, we show that this "adhesive" region of the protein binds directly to fibronectin. Fibronectin fusion proteins corresponding to the various functional domains were used to define the second heparin-binding domain as the ADAM13 binding site. Mutation of the syndecan-binding site (PPRR 3 PPTM) within this domain abolishes binding of the recombinant disintegrin and cysteine-rich domains of ADAM13. We further show that the adhesive disintegrin and cysteine-rich domain of ADAM13 can promote cell adhesion via ␤ 1 integrins. This adhesion requires integrin activation and can be prevented by antibodies to the cysteine-rich domain of ADAM13 and ␤ 1 integrin. Finally, wild type, but not the E/A mutant of ADAM13 metalloprotease domain, can be shed from the cell surface, releasing the metalloprotease domain associated with the disintegrin and cysteinerich domains. This suggests that ADAM13 shedding may involve its own metalloprotease activity and that the released protease may interact with both integrins and extracellular matrix proteins. ADAMs1 are transmembrane glycoproteins that contain a disintegrin and a metalloprotease domain (1). To date, 33 ADAMs have been identified in worms, insects, amphibians, and mammals (2). They can be classified in two major subgroups depending on the presence or absence of a putative active site (HEXGHXXGXHD) in the metalloprotease domain (3). ADAM proteins have been implicated in many biological processes including fertilization (4, 5), myogenesis (6), neurogenesis (7), axon extension and pathfinding (8), and neural crest cell migration (9). Although the number of potential functions involving ADAMs increases, the function of individual ADAM protein domains remains poorly understood.The ADAM metalloprotease domain appears to be involved mostly in shedding cell surface molecules (10). Among these molecules are growth factors (tumor necrosis factor-␣, transforming growth factor-␣, heparin binding-epidermal growth factor), other signaling molecules (Notch, Delta, ephrin), and cell adhesion molecules (selectin, L1-Cam). These cleavages allow cells to send and/or receive a signal as well as change their adhesion to the surrounding cells or substrate. Recently, ADAM13 was shown to cleave fibronectin, one of the major components of the extracellular matrix (ECM) that lies in all cell migration pathways (9).The disintegrin and cysteine-rich domains compose the adhesive part of the ADAM protein. A small loop within the disintegrin domain, known as the disintegrin loop, can bind to several receptors of the integrin family (11). The best example of this is the interaction of ADAM2 and -3 on the mature mouse sperm with the ␣ 6 ␤...

show abstract

“…It will be important to consider whether ␣ 5 ␤ 1 can distinguish soluble Fg, immobilized Fg, or fibrin and plasmic degradation products of Fg/fibrin. Precedence exists for differential recognition of various forms of Fg and other ligands by integrins (54). Thus, the availability of specific receptors on the luminal surface, the activation states Fibrinogen Binding to ␣ 5 ␤ 1 on HUVECof the EC and these receptors, the occupancy of these receptors by competing ligands, and the nature of the ligands may determine which receptors will mediate Fg recognition by HUVEC.…”

Section: Discussionmentioning

confidence: 99%

Fibrinogen Is a Ligand for Integrin α5β1 on Endothelial Cells

Suehiro

Gailit

Plow

1997

Journal of Biological Chemistry

Self Cite

165

129

View full text Add to dashboard Cite

Previous studies have shown that fibrinogen can associate with endothelial cells via an Arg-Gly-Asp (RGD) recognition specificity. In the present study, we have characterized the specificity of fibrinogen binding to endothelial cells under different cation conditions. Fibrinogen binding to suspended endothelial cells was selectively supported by Mn 2؉ and was suppressed by Ca 2؉ . The Mn 2؉ -supported interaction was completely inhibited by RGD peptides but not by ␣ v ␤ 3 blocking monoclonal antibodies. In contrast, the interaction was completely blocked by two ␣ 5 ␤ 1 monoclonal antibodies. This interaction was not mediated by fibronectin bound to the integrin; could be demonstrated with purified ␣ 5 ␤ 1 ; and also was observed with a second ␣ 5 ␤ 1 -bearing cell type, platelets. The binding of fibrinogen to ␣ 5 ␤ 1 on endothelial cells in the presence of Mn 2؉ was time-dependent, specific, saturable, and of high affinity (K d ‫؍‬ 65 nM). By employing anti-peptide monoclonal antibodies, the carboxyl-terminal RGD sequence at A␣ 572-574 was implicated in fibrinogen recognition by ␣ 5 ␤ 1 . Two circumstances were identified in which ␣ 5 ␤ 1 interacted with fibrinogen in the presence of Ca 2؉ : when the receptor was activated with monoclonal antibody (8A2) or when the fibrinogen was presented as an immobilized substratum. These results identify fibrinogen as a ligand for ␣ 5 ␤ 1 on endothelial and other cells, an interaction which may have broad biological implications.The luminal surface of endothelial cells (EC) 1 is continuously exposed to a high concentration of plasma fibrinogen (Fg). Disruption of the endothelium results in local thrombus formation, and Fg/fibrin accumulates at such sites of vascular injury. Based upon these proximal relationships, the molecular mechanisms and functional consequences of the interaction of Fg with EC have been topics of considerable interest and investigation (1-6). Indeed, it has been shown that Fg can induce EC attachment, spreading, and migration (1, 7) and can support an angiogenic response (8). Several distinct receptors have been implicated in mediating Fg binding to EC. These include ␣ v ␤ 3 (9, 10), a member of the integrin family of cell adhesion receptors, as well as several non-integrin binding sites (6, 11). Transglutaminase-mediated Fg cross-linking to EC also has been demonstrated (12).␣ v ␤ 3 interacts with Fg via an Arg-Gly-Asp (RGD) recognition specificity; i.e. RGD-containing peptides block Fg binding to this receptor (13). This tripeptide sequence is recognized not only by ␣ v ␤ 3 but also by several other integrins (14 -16), including ␣ 5 ␤ 1 , which serves as a fibronectin (Fn) receptor on EC and many other cell types (17)(18)(19)(20). Fg contains two RGD sequences within its A␣-chain: RGDF at A␣ 95-98 and RGDS at A␣ 572-575. Previous studies have shown that EC adhesion to immobilized Fg is blocked by a monoclonal antibody (mAb) to the carboxyl-terminal peptide containing RGD sequence at A␣ 572-574 (3). Moreover, this adhesion was blocked by mAbs against ␣...

show abstract

Proteolysis Regulates Exposure of the IIICS-1 Adhesive Sequence in Plasma Fibronectin

Cited by 36 publications

References 59 publications

Granzyme B degrades extracellular matrix and contributes to delayed wound closure in apolipoprotein E knockout mice

Granzyme B degrades extracellular matrix and contributes to delayed wound closure in apolipoprotein E knockout mice

ADAM13 Disintegrin and Cysteine-rich Domains Bind to the Second Heparin-binding Domain of Fibronectin

Fibrinogen Is a Ligand for Integrin α5β1 on Endothelial Cells

Contact Info

Product

Resources

About