177 Distinguishing the specificities of closely related proteases: Role of P3 in substrate and inhibitor discrimination between tissue type plasminogen activator and urokinase

Ke, Song-Hua; Coobms, Gary S.; Tachias, Kathy; Navre, Marc; Corey, David R.; Madison, Edwin L.

doi:10.1016/s0268-9499(97)80292-5

Cited by 10 publications

(13 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our analysis also demonstrated that the conserved tribasic -R 254 KSK-site in PDGF-DD was required for effective activation, even though the actual processing by uPA is likely confined to -R 249 . This is further supported by a previous report suggesting a preference for glycine at the P2 subsite of tPA and uPA substrates (Ke et al, 1997). Owing to its small size, glycine is expected to render the basic cleavage site more exposed to activating proteases and accordingly, this may also explain why removal of the bulky arginine side chain in the R247A mutation enhanced PDGF-DD activation.…”

Section: Discussionsupporting

confidence: 83%

The uPA/uPAR system regulates the bioavailability of PDGF-DD: implications for tumour growth

Ehnman

Fredriksson

et al. 2008

Oncogene

View full text Add to dashboard Cite

Members of the platelet-derived growth factor (PDGF) family are mitogens for cells of mesenchymal origin and have important functions during embryonic development, blood vessel maturation, fibrotic diseases and cancer. In contrast to the two classical PDGFs, the novel and less well-characterized members, PDGF-CC and PDGF-DD, are latent factors that need to be processed extracellularly by activating proteases, before they can mediate PDGF receptor activation. Here, we elucidate the structural requirements for urokinase plasminogen activator (uPA)-mediated activation of PDGF-DD, as well as the intricate interplay with uPA receptor (uPAR) signalling. Furthermore, we show that activated PDGF-DD, in comparison to latent, more potently transforms NIH/3T3 cells in vitro. Conversely, xenograft studies in nude mice demonstrate that cells expressing latent PDGF-DD are more tumorigenic than those expressing activated PDGF-DD. These findings imply that a fine-tuned proteolytic activation, in the local milieu, controls PDGF-DD bioavailability. Moreover, we suggest that proteolytic activation of PDGF-DD reveals a retention motif mediating interactions with pericellular components. Our proposed mechanism, where uPA not only generates active PDGF-DD, but also regulates its spatial distribution, provides novel insights into the biological function of PDGF-DD.

show abstract

Section: Discussionsupporting

confidence: 83%

The uPA/uPAR system regulates the bioavailability of PDGF-DD: implications for tumour growth

Ehnman

Fredriksson

et al. 2008

Oncogene

View full text Add to dashboard Cite

show abstract

“…However, there was sequence similarity between the R247/R249 region of PDGF D and the preferred cleavage site of uPA identified by phage library display and confirmed in an engineered protein (Fig. 7A) (6,14,15). To determine whether this putative uPA cleavage site of PDGF D is critical for the uPA-mediated proteolytic cleavage of PDGF D, R247 and R249 were both mutated to alanines by site-directed mutagenesis and then the mutant PDGF D R247,249A was inserted into the vaccinia virus expression vector for further analysis (Fig.…”

Section: Resultsmentioning

confidence: 87%

Platelet-Derived Growth Factor D Is Activated by Urokinase Plasminogen Activator in Prostate Carcinoma Cells

Ustach

Kim

2005

Molecular and Cellular Biology

104

View full text Add to dashboard Cite

Platelet-derived growth factors (PDGFs) regulate a diverse array of cellular processes, including cell proliferation, transformation, migration, survival, and apoptosis of mesenchymal cells, in development as well as during pathogenesis (reviewed in references 31 and 42). For over 2 decades, PDGFs were thought to exist as the homodimers PDGF AA and BB or the heterodimer PDGF AB. These PDGF dimers are processed intracellularly and secreted as active dimers that readily activate PDGF receptors (PDGFRs). Recently, two new PDGF ligands (PDGF CC and DD) were discovered that have a unique two-domain structure with an N-terminal complement subcomponent C1r/C1s, Uegf, Bmp1 (CUB) domain and a C-terminal PDGF/vascular endothelial growth factor domain. PDGF CC and DD are secreted as full-length, latent dimers, and the proteolytic removal of the CUB domain is required for the growth factor domain of PDGF CC or DD to activate the PDGF receptors (4,20,21).PDGFs exert their biological functions through the activation of dimeric receptors made up of two structurally similar protein-tyrosine kinase receptor subunits (␣␣-, ␣␤-, or ␤␤-

show abstract

“…In Silico Prediction of Cleavage Sites-Specific cleavage sites for both uPA and tPA were confirmed with phage substrate libraries (43)(44)(45)(46)(47). The consensus motif in substrates for proteolytic cleavage by urokinase is GR2(SϾN/K/R)(AϾ ϾS) from P2 to P2Ј.…”

Section: Methodsmentioning

confidence: 87%

“…A series of classic studies on the specificity of uPA substrates revealed a consensus cleavage motif, GR2(SϾN/K/R)(AϾ ϾS) from P2 to P2Ј (43,44). We further combined in silico prediction and immunoblotting assays to narrow down the cleavage site.…”

Section: Proteolysis Of Deletion Mutants Missing Consensus Motifs Formentioning

confidence: 99%

Proteolytic Regulation of Epithelial Sodium Channels by Urokinase Plasminogen Activator

Zhao²,

Komissarov³

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Depressed fibrinolysis and edema concurrently exist in edematous injury. Results: Divergent regulation of ENaC by urokinase and tPA was observed. Both the catalytic domain of urokinase and cleavage sites in ENaC were identified. Conclusion: Urokinase activates ENaC through catalytic activity-dependent proteolytic modification of ␥ subunit. Significance: Activation of ENaC by urokinase but not tPA provides a novel mechanism for the alleviation of lung edema and pleural effusion.

show abstract

177 Distinguishing the specificities of closely related proteases: Role of P3 in substrate and inhibitor discrimination between tissue type plasminogen activator and urokinase

Cited by 10 publications

References 0 publications

The uPA/uPAR system regulates the bioavailability of PDGF-DD: implications for tumour growth

The uPA/uPAR system regulates the bioavailability of PDGF-DD: implications for tumour growth

Platelet-Derived Growth Factor D Is Activated by Urokinase Plasminogen Activator in Prostate Carcinoma Cells

Proteolytic Regulation of Epithelial Sodium Channels by Urokinase Plasminogen Activator

Contact Info

Product

Resources

About