Utilization of biotinylated diphenyl phosphonates for disclosure of serine proteases

Hawthorne, Susan; Hamilton, Robert A.; Walker, Brian

doi:10.1016/j.ab.2003.12.002

Cited by 21 publications

(18 citation statements)

References 8 publications

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“…To detect trypsin-like enzymes in the cockroach extracts, we employed a strategy described previously for the identification of serine proteinases using biotinylated diphenyl phosphonate probes (27). On the basis of the use of probes with proline-lysine or asparagine-lysine sequences in the P2 and P1 enzyme target positions (28), we elected to synthesize the biotinylated probe with the PK sequence as the tryptic target (so-called compound 4, biotin-linker-Pro-Lys-diphenyl phosphonate, Bio-PK-DPP4).…”

Section: Serine Proteinase Activity-based Probementioning

confidence: 99%

Mucosal Allergic Sensitization to Cockroach Allergens Is Dependent on Proteinase Activity and Proteinase-Activated Receptor-2 Activation

Arizmendi

Abel

Mihara

et al. 2011

The Journal of Immunology

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We have shown that proteinase-activated receptor-2 (PAR2) activation in the airways leads to allergic sensitization to concomitantly inhaled Ags, thus implicating PAR2 in the pathogenesis of asthma. Many aeroallergens with proteinase activity activate PAR2. To study the role of PAR2 in allergic sensitization to aeroallergens, we developed a murine model of mucosal sensitization to cockroach proteins. We hypothesized that PAR2 activation in the airways by natural allergens with serine proteinase activity plays an important role in allergic sensitization. Cockroach extract (CE) was administered to BALB/c mice intranasally on five consecutive days (sensitization phase) and a week later for four more days (challenge phase). Airway hyperresponsiveness (AHR) and allergic airway inflammation were assessed after the last challenge. To study the role of PAR2, mice were exposed intranasally to a receptor-blocking anti-PAR2 Ab before each administration of CE during the sensitization phase. Mucosal exposure to CE induced eosinophilic airway inflammation, AHR, and cockroach-specific IgG1. Heat-inactivated or soybean trypsin inhibitor-treated CE failed to induce these effects, indicating that proteinase activity plays an important role. The use of an anti-PAR2 blocking Ab during the sensitization phase completely inhibited airway inflammation and also decreased AHR and the production of cockroach-specific IgG1. PAR2 activation by CE acts as an adjuvant for allergic sensitization even in the absence of functional TLR4. We conclude that CE induces PAR2-dependent allergic airway sensitization in a mouse model of allergic airway inflammation. PAR2 activation may be a general mechanism used by aeroallergens to induce allergic sensitization.

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Section: Serine Proteinase Activity-based Probementioning

confidence: 99%

Mucosal Allergic Sensitization to Cockroach Allergens Is Dependent on Proteinase Activity and Proteinase-Activated Receptor-2 Activation

Arizmendi

Abel

Mihara

et al. 2011

The Journal of Immunology

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“…This approach, targeting secreted enzymes, circumvents problems encountered with the use of biotin-tagged reagents for cell-based analyses, because in cells or tissues the high levels of endogenous background biotin-containing proteins can be problematic (Sadaghiani et al, 2007;Fonović et al, 2007). The ABP we used was designed in accordance with the strategy for tagging serine proteinases with a biotinylated diphenylphosphonate probe (Hawthorne et al, 2004). The phosphonate reactive group (which alkylates the serine residue within the active site of any serine proteinase) is preceded by a prolinelysine sequence in the P2/P1 enzyme target site to restrict the probe specificity to trypsin-like serine proteinases ( Figure 2A).…”

Section: Activity-based Probe Elisamentioning

confidence: 99%

Kallikrein-related peptidases: proteolysis and signaling in cancer, the new frontier

Οικονομοπούλου¹,

Diamandis²,

Hollenberg³

2010

Biological Chemistry

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The exact mechanism(s) by which kallikrein-related peptidases (KLKs) function, their levels of activity and their potential endogenous targets in vivo have only recently begun to be revealed. Our group and others have shown that KLKs can have hormonal properties by signaling via proteinase-activated receptors (PARs), a family of G-protein-coupled receptors. Signals by PAR(1), PAR(2), and PAR(4) can regulate calcium release or mitogen-activated protein kinase activation and lead to platelet aggregation, vascular relaxation, cell proliferation, cytokine release, and inflammation. We have further documented the presence of active KLK6 and 10 (by activity-based ELISA or proteomics) and the presence of proteinase inhibitors, such as alpha(1)-antitrypsin, in cancer-derived fluids. We suggest that tumors and inflamed tissues can release active KLKs, which are under tight regulation by proteinase inhibitors. These enzymes can potentially control cell/tissue behavior by regulating PAR activation in specific settings and disease stages.

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“…To label rISP2 with an activity-based probe (ABP), we used a strategy previously described for the identification of serine proteinases using biotinylated diphenyl phosphonate probes (Hawthorne et al 2004). Based on the synthesis of probes with proline-lysine or asparagine-lysine sequences in the P2 and P1 enzyme target positions (Pan et al 2006), we elected to use the biotinylated probe with the PK sequence as the tryptic target (socalled, compound 4, Bio-PK-DPP 4).…”

Section: Activity-based Serine Proteinase Probe Labelling Of Isp2mentioning

confidence: 99%

Implantation serine proteinase 2 is a monomeric enzyme with mixed serine proteolytic activity and can silence signalling via proteinase activated receptors

Sharma

Fahr

Renaux

et al. 2013

Biochem. Cell Biol.

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Implantation serine proteinase 2 (ISP2), a S1 family serine proteinase, is known for its role in the critical processes of embryo hatching and implantation in the mouse uterus. Native implantation serine proteinases (ISPs) are co-expressed and co-exist as heterodimers in uterine and blastocyst tissues. The ISP1-ISP2 enzyme complex shows trypsin-like substrate specificity. In contrast, we found that ISP2, isolated as a 34 kDa monomer from a Pichia pastoris expression system, exhibited a mixed serine proteolytic substrate specificity, as determined by a phage display peptide cleavage approach and verified by the in vitro cleavage of synthetic peptides. Based upon the peptide sequence substrate selectivity, a database search identified many potential ISP2 targets of physiological relevance, including the proteinase activated receptor 2 (PAR2). The in vitro cleavage studies with PAR2-derived peptides confirmed the mixed substrate specificity of ISP2. Treatment of cell lines expressing proteinase-activated receptors (PARs) 1, 2, and 4 with ISP2 prevented receptor activation by either thrombin (PARs 1 and 4) or trypsin (PAR2). The disarming and silencing of PARs by ISP2 may play a role in successful embryo implantation.

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Utilization of biotinylated diphenyl phosphonates for disclosure of serine proteases

Cited by 21 publications

References 8 publications

Mucosal Allergic Sensitization to Cockroach Allergens Is Dependent on Proteinase Activity and Proteinase-Activated Receptor-2 Activation

Mucosal Allergic Sensitization to Cockroach Allergens Is Dependent on Proteinase Activity and Proteinase-Activated Receptor-2 Activation

Kallikrein-related peptidases: proteolysis and signaling in cancer, the new frontier

Implantation serine proteinase 2 is a monomeric enzyme with mixed serine proteolytic activity and can silence signalling via proteinase activated receptors

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