Mast cells protect from post-traumatic spinal cord damage in mice by degrading inflammation-associated cytokines via mouse mast cell protease 4

Five highly soluble, chymotrypsin-like, neutral serine proteases, with molecular masses in the range 30-33 kDa, were isolated from Trichinella spiralis-infected mouse small intestine. These enzymes were closely related antigenically on Western blotting and by Ouchterlony double diffusion using a polyclonal, cross-absorbed, sheep antibody raised against mouse mast cell protease-1 (MMCP-1) and on the basis of N-terminal amino acid sequence analysis, were identified as variant forms of MMCP-1. Substrate and inhibitor analysis confirmed that the five variants (MMCP-1 A-E) had similar characteristics, although highly significant (P = 0.025 to P < 0.0001) variations in Km and kcat, were detected. Against human alpha 1-proteinase inhibitor the Ki for MMCP-1C (45 pM) was significantly (P < 0.0001) greater than those for the other proteases (0.76-2.2 pM). The differences in electrophoretic mobility are probably a result of variable glycosylation, since removal of N-linked carbohydrate produced a polypeptide of approx. 28 kDa in each case which was, like the native enzyme, immunoreactive on Western blotting. A much less soluble 28 kDa enzyme was isolated from serosal mast cells and identified as MMCP-4 by N-terminal amino acid sequencing. Like MMCP-1 it has chymotrypsin-like substrate specificities with activity at neutral pH. However, it was antigenically distinct from MMCP-1 and, using sheep anti-MMCP-1, was not detected on Western blotting or by Ouchterlony double diffusion, e.l.i.s.a. or immunohistochemistry. This last technique established that the MMCP-1 variants were uniquely present in enteric mast cells, thereby providing a highly selective means of distinguishing the mucosal and connective tissue mast cell subsets in the mouse.

Download Full-text

Mouse mast cell protease 4 suppresses scar formation after traumatic spinal cord injury

Scientific Reports ◽

10.1038/s41598-019-39551-1 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 5

Author(s):

Tim Vangansewinkel ◽

Stefanie Lemmens ◽

Nathalie Geurts ◽

Kirsten Quanten ◽

Dearbhaile Dooley ◽

...

Keyword(s):

Spinal Cord ◽

Spinal Cord Injury ◽

Mast Cell ◽

Scar Formation ◽

Traumatic Spinal Cord Injury ◽

Mast Cell Protease ◽

Cord Injury ◽

Mouse Mast Cell

Download Full-text

Basophils preferentially express mouse mast cell protease 11 among the mast cell tryptase family in contrast to mast cells

Journal of Leukocyte Biology ◽

10.1189/jlb.0609400 ◽

2009 ◽

Vol 86 (6) ◽

pp. 1417-1425 ◽

Cited By ~ 77

Author(s):

Tsukasa Ugajin ◽

Toshiyuki Kojima ◽

Kaori Mukai ◽

Kazushige Obata ◽

Yohei Kawano ◽

...

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Mast Cell Tryptase ◽

Mast Cell Protease ◽

Mouse Mast Cell

Download Full-text

Cloning of the cDNA and gene of mouse mast cell protease-6. Transcription by progenitor mast cells and mast cells of the connective tissue subclass

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)67871-3 ◽

1991 ◽

Vol 266 (6) ◽

pp. 3847-3853

Author(s):

D S Reynolds ◽

D S Gurley ◽

K F Austen ◽

W E Serafin

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Connective Tissue ◽

Mast Cell Protease ◽

Mouse Mast Cell

Download Full-text

Requirement of c-jun transcription factor on the mouse mast cell protease-6 expression in the mast cells

Archives of Biochemistry and Biophysics ◽

10.1016/j.abb.2004.07.027 ◽

2004 ◽

Vol 431 (1) ◽

pp. 71-78 ◽

Cited By ~ 8

Author(s):

Dae-Ki Kim ◽

Young-Mi Lee

Keyword(s):

Transcription Factor ◽

Mast Cell ◽

Mast Cells ◽

Mast Cell Protease ◽

Mouse Mast Cell

Download Full-text

Mouse bone marrow-derived mast cells (mBMMC) obtained in vitro from mice that are mast cell-deficient in vivo express the same panel of granule proteases as mBMMC and serosal mast cells from their normal littermates.

Journal of Experimental Medicine ◽

10.1084/jem.180.1.67 ◽

1994 ◽

Vol 180 (1) ◽

pp. 67-73 ◽

Cited By ~ 26

Author(s):

K K Eklund ◽

N Ghildyal ◽

K F Austen ◽

D S Friend ◽

V Schiller ◽

...

Keyword(s):

Bone Marrow ◽

Mast Cell ◽

Steady State ◽

Mast Cells ◽

Induced Expression ◽

Mast Cell Protease ◽

Steady State Levels ◽

Mouse Mast Cell

The ear, skin, and purified serosal mast cells of WBB6F1/J-(+/+) (WB-(+/+)) and WCB6F1/J-(+/+) (WC-(+/+)) mice contain high steady-state levels of the transcripts that encode mouse mast cell protease (mMCP) 2, mMCP-4, mMCP-5, mMCP-6, and mouse mast cell carboxypeptidase A (mMC-CPA). In contrast, no mast cell protease transcripts are present in abundance in the ear and skin of WBB6F1/J-W/Wv (W/Wv) and WCB6F1/J-Sl/Sld (Sl/Sld) mice which are mast cell-deficient in vivo due to defects in their c-kit and c-kit ligand genes, respectively. We now report that the immature bone marrow-derived mast cells (mBMMC) obtained in vitro with recombinant interleukin 3 (rIL-3) or WEHI-3 cell conditioned medium from WB-(+/+), WC-(+/+), W/Wv, and Sl/Sld mice all contain high steady-state levels of the mMCP-2, mMCP-4, mMCP-5, mMCP-6, and mMC-CPA transcripts. As assessed immunohistochemically, mMCP-2 protein and mMCP-5 protein are also present in the granules of mBMMC from WB-(+/+), WC-(+/+), and W/Wv mice. That Sl/Sld and W/Wv mBMMC contain high steady-state levels of five granule protease transcripts expressed by the mature serosal, ear, and skin mast cells of their normal +/+ littermates suggests that c-kit-mediated signal transduction is not essential for inducing transcription of these protease genes. Because rIL-4 inhibits the rIL-10-induced expression of mMCP-1 and mMCP-2 in BALB/cJ mBMMC, the ability of rIL-4 to influence protease mRNA levels in WC-(+/+) mBMMC and W/Wv mBMMC was investigated. Although rIL-10 induced expression of the mMCP-1 transcript in WC-(+/+) and W/Wv mBMMC, rIL-4 was not able to suppress the steady-state levels of the mMCP-1 transcript or any other protease transcript in these cultured mast cells. Thus, not only do BALB/cJ mBMMC express fewer granule proteases than mBMMC from mast cell-deficient strains and their normal littermates but the subsequent induction of late-expressed proteases in BALB/cJ mBMMC is more tightly regulated by IL-3 and IL-4.

Download Full-text

Faculty Opinions recommendation of Basophils preferentially express mouse Mast Cell Protease 11 among the mast cell tryptase family in contrast to mast cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1163808.625608 ◽

2009 ◽

Author(s):

Richard L Stevens

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Mast Cell Tryptase ◽

Mast Cell Protease ◽

Mouse Mast Cell

Download Full-text

Abnormal Expression of Mouse Mast Cell Protease 5 Gene in Cultured Mast Cells Derived From Mutant mi/mi Mice

Blood ◽

10.1182/blood.v90.8.3057 ◽

1997 ◽

Vol 90 (8) ◽

pp. 3057-3066 ◽

Cited By ~ 73

Author(s):

Eiichi Morii ◽

Tomoko Jippo ◽

Tohru Tsujimura ◽

Koji Hashimoto ◽

Dae-Ki Kim ◽

...

Keyword(s):

Mast Cell ◽

Transcription Factors ◽

Mast Cells ◽

Promoter Region ◽

Leucine Zipper ◽

Consensus Sequence ◽

Mast Cell Protease ◽

Helix Loop Helix ◽

Regulation Mechanisms ◽

Mouse Mast Cell

Abstract Mast cells contain a lot of mast cell-specific proteases. We have reported that the expression of mouse mast cell protease 6 (MMCP-6) is remarkably reduced in both cultured mast cells (CMCs) and skin mast cells of mi/mi mutant mice. In the present study, we found that the expression of MMCP-5 was reduced in CMCs but not in skin mast cells of mi/mi mice, and we compared the regulation mechanisms of MMCP-5 with those of MMCP-6. The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper (bHLH-Zip) protein family of transcription factors (hereafter called MITF ). The consensus sequence recognized and bound by bHLH-Zip transcription factors is CANNTG. The overexpression of the normal (+) MITF but not of mi-MITF normalized the poor expression of the MMCP-5 gene in mi/mi CMCs, indicating the involvement of +-MITF in transactivation of the MMCP-5 gene. Although +-MITF directly bound CANNTG motifs in the promoter region of the MMCP-6 gene and transactivated it, the binding of +-MITF to the CAGTTG motif in the promoter region of the MMCP-5 gene was not detectable. The +-MITF appeared to regulate the transactivation of the MMCP-5 gene indirectly. Moreover, addition of stem cell factor to the medium normalized the expression of the MMCP-5 but not of the MMCP-6 gene in mi/mi CMCs. Despite the significant reduction of both MMCP-5 and MMCP-6 expressions in mi/mi CMCs, their regulation mechanisms appeared to be different.

Download Full-text