Chymotrypsin- and trypsin-type serine proteases in rat mast cells: Properties and functions

Kido, Hiroshi; Fukusen, Naomi; Katunuma, Nobuhiko

doi:10.1016/0003-9861(85)90709-x

Cited by 121 publications

(69 citation statements)

References 19 publications

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“…To provide evidence supporting a functionally active RVCH, A10, and aortic and pulmonary artery SMCs from 8-week-old Sprague-Dawley, WKY, and SHR rats (Charles River Laboratories) were used. Samples were prepared according to a paper published previously (44). The vesicles were extracted from SMCs and resuspended in 20 mM Tris-HCl buffer, pH 8.0.…”

Section: Figurementioning

confidence: 99%

A novel vascular smooth muscle chymase is upregulated in hypertensive rats

Guo¹,

Ju²,

Leung³

et al. 2001

J. Clin. Invest.

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Section: Figurementioning

confidence: 99%

A novel vascular smooth muscle chymase is upregulated in hypertensive rats

Guo¹,

Ju²,

Leung³

et al. 2001

J. Clin. Invest.

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“…The SMC were disrupted in 20 mM Tris⅐HCl buffer (pH 8.0) containing 0.5 mM EDTA, 1 g͞ml leupeptin, and 0.5 g͞ml pepstatin A. A synthetic fluorescent substrate, Suc-Leu-Leu-Val-Tyr-7-amino-4-methylcoumarin, was used to measure chymase activity in the lysates, defined as fluorescent units͞min per mg protein (22). To archive conditional and targeted expression of RVCH in SMC of transgenic mice, we used the Tet-Off binary system.…”

Section: Immunoprecipitation Of Rvchmentioning

confidence: 99%

Conditional and targeted overexpression of vascular chymase causes hypertension in transgenic mice

Gros²,

You³

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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We cloned a rat vascular chymase (RVCH) from smooth muscle cells (SMCs) that converts angiotensin I to II and is up-regulated in SMC from spontaneously hypertensive vs. normotensive rats. To determine whether increased activity of RVCH is sufficient to cause hypertension, transgenic mice were generated with targeted conditional expression of RVCH to SMC, with the use of the tetracyclinecontrolled transactivator (tTA). We confirmed conditional expression of RVCH by mRNA, protein, and chymase activity in the absence, but not in the presence, of dietary doxycycline. The systolic blood pressure (mmHg), measured by carotid artery cannulation at 10 -12 weeks of age, was higher in tTA؉͞RVCH؉ mice than in nonbinary transgenic littermates (136 ؎ 4 vs. 109 ؎ 3) (P < 0.05), as were the diastolic and mean pressures. Hypertension was completely reversed by doxycycline, suggesting a causal link with chymase expression. Medial thickening of mesenteric arteries from tTA؉͞RVCH؉ mice vs. littermates (0.82 ؎ 0.1 vs. 0.42 ؎ 0.02) (P < 0.05) was associated with increased SMC proliferation, as judged by positive immunoreactivity, with the use of an antibody to the proliferating cell nuclear antigen. These structural changes were prevented by doxycycline. Perfusion myography of mesenteric arteries from tTA؉͞RVCH؉ mice also revealed increased vasoconstriction in response to phenylephrine and impaired metacholine-induced vasodilatation when compared with littermate controls or with the doxycyline-treated group. Our studies suggest that up-regulation of this vascular chymase is sufficient to cause a hypertensive arteriopathy, and that RVCH may be a candidate gene and a therapeutic target in patients with high blood pressure.smooth muscle cells C hymases are serine proteinases that were thought to be produced exclusively by mast cells in blood vessels and the myocardium. These enzymes generate angiotensin (Ang) II in large and resistance vessels (1-3) and in the heart (4) and also can convert big-endothelin-1 to endothelin-1 in vitro (5) and inactivate the vasodilator bradykinin (6). That chymase-dependent Ang II formation can occur in the intact animal is evident from studies in the baboon, where it was shown that Ang-converting enzyme inhibitors could not prevent the hemodynamic response to [Pro11, Da1a12]Ang I, which produces Ang II upon incubation with chymase (7). In addition to blood pressure regulation, both Ang II and endothelin-1 are involved in the stimulation of vascular smooth muscle cell (SMC) growth and proliferation, vascular remodeling, and cardiac hypertrophy (8, 9). Chymases also can influence structural remodeling through their ability to degrade extracellular matrix proteins, including fibronectin and collagen type IV, possibly through activation of matrix metalloproteinases (MMPs) (MMP-1, MMP-3, and MMP-9) (10-12). Chymases also stimulate collagen fibirillogenesis by cleavage of type I procollagen (13).Several studies have provided compelling data to suggest that chymases could play a central role in cardiovascular di...

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“…Rat CTMC are known to contain at least three different proteolytic enzymes ; a carboxypeptidase A-like enzyme (30), a trypsin-like enzyme (11), and a chymotrypsin-like enzyme (30). Only a chymotrypsin-like protease and none of the others has been found to date in MMC (30).…”

Section: Mast Cell Proteasesmentioning

confidence: 99%