Tryptase in Rat Mast Cells: Properties and Inhibition by Various Inhibitors in Comparison with Chymase

Muramatu, Mutumi; Itoh, Tadashi; Takei, Masao; Endo, Kenji

doi:10.1515/bchm3.1988.369.2.617

Cited by 14 publications

(5 citation statements)

References 15 publications

(5 reference statements)

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“…Two of the other basic proteins (PB6, 29 kDa and PB7, 28 kDa) also bound [3H]DFP, suggesting that they too were serine proteases. Although their identity remains unknown, tryptic activities that could be inhibited by DFP have been found in rat PMC [9,22]. In an attempt to determine whether PB6 or PB7 might be a subunit of rat PMC tryptase [9, 221, sonicated rat PMC were divided into two aliquots, one of which was incubated with 10 ph4 leupeptin [9] for 1 h at 37 "C, and both aliquots were then incubated with [3H]DFP for either 15 min or 2 h. Since leupeptin has been shown to inhibit rat PMC tryptase but not RMCP I [9,22], it was anticipated that time-dependent differences in DFP binding between PB8 (RMCP I) and PB6 and/or PB7 would be found.…”

Section: Discussionmentioning

confidence: 99%

Mast cell heterogeneity: two‐dimensional gel electrophoretic analyses of rat peritoneal and intestinal mucosal mast cell

et al. 1990

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Peritoneal mast cells (PMC) and intestinal mucosal mast cells (IMMC) were purified from rats infected with the nematode Nippostrongylus brasiliensis. Overall protein constituents of both mast cell subtypes were analyzed by two-dimensional gel electrophoresis using either nonequilibrium pH gradient electrophoresis (NEPHGE) or isoelectric focusing (IEF) in the first dimension and SDS-PAGE (10%) in the second dimension followed by silver staining. PMC had seven dominant basic proteins (PB2-8; pI 9-9.5) with estimated molecular masses of 26 to 37 kDa, as well as 80 to 90 neutral or acidic proteins, most of which had pI 6 to 7.5 and estimated molecular masses of 20 to 100 kDa. All the basic proteins were granule-associated. Three basic proteins, PB6 (29 kDa), PB7 (28 kDa) and PB8 (RMCP I, 26 kDa), bound [3H]diisopropyl fluorophosphate (DFP), suggesting that they are serine proteases. However, only PB8 was reactive with antibodies to RMCP I. Another basic component (less than 14 kDa), perhaps a degradation product of PB6, PB7 or PB8, also bound [3H]DFP. By comparison, IMMC possessed nine basic proteins (IB1-9) and, in general, they were more acidic (pI about 8.5-9) than those of PMC. Four major basic proteins (IB6-9) were all 24 kDa but were slightly different in isoelectric points. These and another 46-kDa basic component (IB2) were reactive with antibodies to RMCP II and bound [3H]DFP. There were no other DFP-binding proteins in IMMC. In spite of remarkable differences between basic granule-associated proteins in PMC and basic proteins in IMMC, spots in the neutral-acidic range were for the most part similar in the two mast cell subsets, although quantitative differences were evident for some spots. Thus, rat mast cell populations from the peritoneal cavity and intestinal mucosa exhibit marked heterogeneity in their protein constituents with basic pI, including in their granule-associated proteins with serine protease activity.

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Section: Discussionmentioning

confidence: 99%

Mast cell heterogeneity: two‐dimensional gel electrophoretic analyses of rat peritoneal and intestinal mucosal mast cell

et al. 1990

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“…The tryptase‐like protease activity determined by the degradation of t‐Boc‐Phe‐Ser‐Arg‐MCA was markedly enhanced by 50–150 mg iodine ml −1 ioxaglate in a concentration‐dependent manner. Although t‐Boc‐Phe‐Ser‐Arg‐MCA is hydrolyzed by other trypsin‐like proteases including trypsin and 73 kDa thiol protease (Molla et al ., 1988), the main proteases included in rat peritoneal mast cells are tryptase, a trypsin‐like protease, and chymase, a chymotrypsin‐like protease (Muramatu et al ., 1988). Moreover, the protease activity was almost completely inhibited by nafamostat mesilate (10 −8 M ).…”

Section: Discussionmentioning

confidence: 99%

“…In (B), ioxaglate (100 mg iodine ml 71 ) was included in the absence or presence of rat peritoneal mast cells (3610 5 cells/well 71 British Journal of Pharmacology vol 138 (5) Tryptase and adverse effects of contrast medium T. Sendo et al in a concentration-dependent manner. Although t-Boc-Phe-Ser-Arg-MCA is hydrolyzed by other trypsin-like proteases including trypsin and 73 kDa thiol protease (Molla et al, 1988), the main proteases included in rat peritoneal mast cells are tryptase, a trypsin-like protease, and chymase, a chymotrypsin-like protease (Muramatu et al, 1988). Moreover, the protease activity was almost completely inhibited by nafamostat mesilate (10 78 M).…”

Section: Discussionmentioning

confidence: 99%

A potent tryptase inhibitor nafamostat mesilate dramatically suppressed pulmonary dysfunction induced in rats by a radiographic contrast medium

Sendo

Itoh

Goromaru

et al. 2003

British J Pharmacology

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1 Intravenous injection of ioxaglate (4 g iodine kg 71), an iodinated radiographic contrast medium, caused a marked protein extravasation, pulmonary oedema and a decrease in the arterial partial oxygen pressure in rats. 2 All of these reactions to ioxaglate were reversed by the pretreatment with gabexate mesilate (10 and 50 mg kg 71, 5 min prior to injection) or nafamostat mesilate (3 and 10 mg kg 71 ), in which the inhibition was complete after injection of nafamostat mesilate (10 mg kg 71 ). 3 Both gabexate mesilate and nafamostat mesilate inhibited the activity of puri®ed human lung tryptase, although the latter compound was far more potent than the former. 4 Ioxaglate enhanced the nafamostat-sensitive protease activity in the extracellular¯uid of rat peritoneal mast cell suspensions. 5 Tryptase enhanced the permeability of protein through the monolayer of cultured human pulmonary arterial endothelial cells. Ioxaglate, when applied in combination with rat peritoneal mast cells, also produced the endothelial barrier dysfunction. These eects of tryptase and ioxaglate were reversed by nafamostat mesilate. 6 Consistent with these ®ndings, immuno¯uorescence morphological analysis revealed that tryptase or ioxaglate in combination with mast cells increased actin stress ®bre formation while decreasing VE-cadherin immunoreactivity. Both of these actions of tryptase and ioxaglate were reversed by nafamostat mesilate. 7 These ®ndings suggest that tryptase liberated from mast cells plays a crucial role in the ioxaglateinduced pulmonary dysfunction. In this respect, nafamostat mesilate may become a useful agent for the cure or prevention of severe adverse reactions to radiographic contrast media. British Journal of Pharmacology (2003) 138, 959 ± 967. doi:10.1038/sj.bjp.0705121 Keywords: Radiographic contrast media; vascular hyperactivity; pulmonary edema; endothelial barrier function; tryptase; actin stress ®bre; VE-cadherin Abbreviations: BSA, bovine serum albumin; HBSS, Hank's balanced salt solution; HPAECs, human pulmonary arterial endothelial cells; PAR-2, proteinase-activated receptor-2; PBS, phosphate buered saline; RCM, radiographic contrast media; t-Boc-Phe-Ser-Arg- IntroductionIn spite of increasing use of radiographic contrast media (RCM), no eective prevention or therapy for the anaphylactoid reactions, including bronchospasm, dyspnea, laryngeal oedema and pulmonary oedema accompanying respiratory distress, induced by the intravascular injection of RCM has yet been established. The pulmonary oedema is a serious lifethreatening adverse event, although the incidence is rare. The RCM-induced pulmonary oedema is a non-cardiogenic type and appears to be related to the increase in pulmonary vascular permeability subsequent to the activation of in¯ammatory cascade or the release of a variety of chemical mediators (Bouachour et al., 1991). Although the precise mechanisms underlying the RCM-induced vascular hyperpermeability or pulmonary dysfunction remain to be clari®ed, several lines of evidence have suggested that ...

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“…This mechanism might be attributed to a common process induced in the histamine release, probably the inhibition of tryptase [6], Muramatu et al [12] suggested the involve ment of a trypsin-like protease in biochemical events during histamine release from rat mast cells. In the present study, we have found that NCO-650 strongly inhibited the cAM P increase induced by antigen, antiIgE and Con A, and the IC50 values were in the micro mole per liter order.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of cAMP Increase by an Anti-Allergic Agent, NCO-650, during Histamine Release

Takei¹,

Matumoto²,

Endo³

et al. 1989

Int Arch Allergy Immunol

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Antigen and concanavalin A (Con A) induced an increase in cAMP and histamine release from rat peritoneal mast cells. In a dose-dependent manner, the compound, NCO-650, significantly inhibited both the initial and secondary increases in cAMP stimulated by antigen, anti-IgE and Con A in rat peritoneal mast cells. IC₅₀ values of NCO-650 for cAMP increase stimulated by antigen, anti-IgE and Con A were 3.8, 3.4 and 2.8 μM, respectively.

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Tryptase in Rat Mast Cells: Properties and Inhibition by Various Inhibitors in Comparison with Chymase

Cited by 14 publications

References 15 publications

Mast cell heterogeneity: two‐dimensional gel electrophoretic analyses of rat peritoneal and intestinal mucosal mast cell

Mast cell heterogeneity: two‐dimensional gel electrophoretic analyses of rat peritoneal and intestinal mucosal mast cell

A potent tryptase inhibitor nafamostat mesilate dramatically suppressed pulmonary dysfunction induced in rats by a radiographic contrast medium

Inhibition of cAMP Increase by an Anti-Allergic Agent, NCO-650, during Histamine Release

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