The substrate degradome of meprin metalloproteases reveals an unexpected proteolytic link between meprin β and ADAM10

Jefferson, Tamara; Keller, Ulrich auf dem; Bellac, Caroline L.; Metz, Verena V.; Broder, Claudia; Hedrich, Jana; Ohler, Anke; Maier, Wladislaw; Magdolen, Viktor; Sterchi, Erwin-Ernst; Bond, Judith S.; Jayakumar, Arumugam; Traupe, Heiko; Chalaris, Athena; Rose-John, Stefan; Pietrzik, Claus U.; Postina, Rolf; Overall, Christopher M.; Becker‐Pauly, Christoph

doi:10.1007/s00018-012-1106-2

Cited by 113 publications

(140 citation statements)

References 87 publications

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“…In addition, data from mass spectrometry revealed two new putative cleavage sites at D 13 [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] ; mass: 2,279.20 Da). This is consistent with the specificity of the active site of meprin-␣, which prefers acidic amino acids at positions near and adjacent to its cleavage site (20,21). Overall, we found four of the most g) from WT and meprin-␣ KO mice.…”

Section: Kh From Meprin-␣ Ko Mice Did Not Release Ac-sdkp From T␤4 Insupporting

confidence: 88%

The anti-inflammatory peptide Ac-SDKP is released from thymosin-β4 by renal meprin-α and prolyl oligopeptidase

Kumar

Nakagawa

Janic

et al. 2016

American Journal of Physiology-Renal Physiology

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The anti-inflammatory peptide Ac-SDKP is released from thymosin-␤4 by renal meprin-␣ and prolyl oligopeptidase. Am J Physiol Renal Physiol 310: F1026 -F1034, 2016. First published March 9, 2016 doi:10.1152/ajprenal.00562.2015.-N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is a natural tetrapeptide with antiinflammatory and antifibrotic properties. Previously, we have shown that prolyl oligopeptidase (POP) is involved in the Ac-SDKP release from thymosin-␤4 (T␤4). However, POP can only hydrolyze peptides shorter than 30 amino acids, and T␤4 is 43 amino acids long. This indicates that before POP hydrolysis takes place, T␤4 is hydrolyzed by another peptidase that releases NH2-terminal intermediate peptide(s) with fewer than 30 amino acids. Our peptidase database search pointed out meprin-␣ metalloprotease as a potential candidate. Therefore, we hypothesized that, prior to POP hydrolysis, T␤4 is hydrolyzed by meprin-␣. In vitro, we found that the incubation of T␤4 with both meprin-␣ and POP released Ac-SDKP, whereas no Ac-SDKP was released when T␤4 was incubated with either meprin-␣ or POP alone. Incubation of T␤4 with rat kidney homogenates significantly released Ac-SDKP, which was blocked by the meprin-␣ inhibitor actinonin. In addition, kidneys from meprin-␣ knockout (KO) mice showed significantly lower basal Ac-SDKP amount, compared with wild-type mice. Kidney homogenates from meprin-␣ KO mice failed to release Ac-SDKP from T␤4. In vivo, we observed that rats treated with the ACE inhibitor captopril increased plasma concentrations of Ac-SDKP, which was inhibited by the coadministration of actinonin (vehicle, 3.1 Ϯ 0.2 nmol/l; captopril, 15.1 Ϯ 0.7 nmol/l; captopril ϩ actinonin, 6.1 Ϯ 0.3 nmol/l; P Ͻ 0.005). Similar results were obtained with urinary Ac-SDKP after actinonin treatment. We conclude that release of Ac-SDKP from T␤4 is mediated by successive hydrolysis involving meprin-␣ and POP.N-acetyl-seryl-aspartyl-lysyl-proline; thymosin-␤4; meprin-␣; prolyl oligopeptidase; angiotensin-converting enzyme THE ANTI-INFLAMMATORY PEPTIDE N-acetyl-seryl-aspartyl-lysylproline (Ac-SDKP) was identified originally in the bone marrow (24). It has also been widely distributed in mammalian organs, plasma, urine, and circulating mononuclear cells (23,39,46). In animal diseases with hypertension, as well as cardiovascular and kidney disease, Ac-SDKP reduces inflammatory cell infiltration and collagen deposition in the heart, kidney, lung, and liver (14,15,33,34,37,44). Ac-SDKP is hydrolyzed by angiotensin-converting enzyme (ACE) and has a short half-life of 4.5 min in the circulation (16). Treatment with ACE inhibitors significantly increased plasma concentration of Ac-SDKP (2), and some of the ACE inhibitors protective effects were reported to be mediated by the increases in Ac-SDKP concentrations (34, 35).Thymosin-␤4 (T␤4) is a major G-actin-sequestering peptide that consists of 43 amino acids. Since it contains the Ac-SDKP sequence in its NH 2 -terminal, T␤4 is considered to be the precursor of 39). Our group has shown p...

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Section: Kh From Meprin-␣ Ko Mice Did Not Release Ac-sdkp From T␤4 Insupporting

confidence: 88%

The anti-inflammatory peptide Ac-SDKP is released from thymosin-β4 by renal meprin-α and prolyl oligopeptidase

Kumar

Nakagawa

Janic

et al. 2016

American Journal of Physiology-Renal Physiology

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“…Interestingly, the cleavage site at Tyr1108/Asp1109 in procollagen α2(I) was previously identified in cell culture using an MS-based approach, terminal amine isotopic labeling of substrates (TAILS) (Fig. 1D) (36). In addition, N-terminal sequencing of the C-propeptide from homotrimeric miniprocollagen α1(I) released after incubation with meprin β confirmed the cleavage site at position Arg1227/Asp1228 (Fig.…”

Section: Resultsmentioning

confidence: 54%

Metalloproteases meprin α and meprin β are C- and N-procollagen proteinases important for collagen assembly and tensile strength

Broder¹,

Arnold

Goff

et al. 2013

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

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102

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Type I fibrillar collagen is the most abundant protein in the human body, crucial for the formation and strength of bones, skin, and tendon. Proteolytic enzymes are essential for initiation of the assembly of collagen fibrils by cleaving off the propeptides. We report that Mep1a −/− and Mep1b −/− mice revealed lower amounts of mature collagen I compared with WT mice and exhibited significantly reduced collagen deposition in skin, along with markedly decreased tissue tensile strength. While exploring the mechanism of this phenotype, we found that cleavage of full-length human procollagen I heterotrimers by either meprin α or meprin β led to the generation of mature collagen molecules that spontaneously assembled into collagen fibrils. Thus, meprin α and meprin β are unique in their ability to process and release both C-and N-propeptides from type I procollagen in vitro and in vivo and contribute to the integrity of connective tissue in skin, with consequent implications for inherited connective tissue disorders.proteolysis | Ehlers-Danlos syndrome | proteomics | fibrosis | connective tissue

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“…Thus, release of the attached GF mucus was dependent on meprin β being secreted from the enterocytes out into the mucus. The metalloproteases ADAM10 and ADAM17 have been shown to be involved in shedding meprin β from the cell surface (21,22). Consequently, we analyzed ADAM10 and 17 mRNA expression levels in whole tissue, but neither of these showed significantly lower levels in GF and are probably therefore not sufficiently altered to explain the lack of meprin β in the GF mucus (Fig.…”

Section: Resultsmentioning

confidence: 99%

Microbial-induced meprin β cleavage in MUC2 mucin and a functional CFTR channel are required to release anchored small intestinal mucus

Schutte¹,

Ermund²,

Becker‐Pauly

et al. 2014

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

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164

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The mucus that covers and protects the epithelium of the intestine is built around its major structural component, the gel-forming MUC2 mucin. The gel-forming mucins have traditionally been assumed to be secreted as nonattached. The colon has a twolayered mucus system where the inner mucus is attached to the epithelium, whereas the small intestine normally has a nonattached mucus. However, the mucus of the small intestine of meprin β-deficient mice was now found to be attached. Meprin β is an endogenous zinc-dependent metalloprotease now shown to cleave the N-terminal region of the MUC2 mucin at two specific sites. When recombinant meprin β was added to the attached mucus of meprin β-deficient mice, the mucus was detached from the epithelium. Similar to meprin β-deficient mice, germ-free mice have attached mucus as they did not shed the membraneanchored meprin β into the luminal mucus. The ileal mucus of cystic fibrosis (CF) mice with a nonfunctional cystic fibrosis transmembrane conductance regulator (CFTR) channel was recently shown to be attached to the epithelium. Addition of recombinant meprin β to CF mucus did not release the mucus, but further addition of bicarbonate rendered the CF mucus normal, suggesting that MUC2 unfolding exposed the meprin β cleavage sites. Mucus is thus secreted attached to the goblet cells and requires an enzyme, meprin β in the small intestine, to be detached and released into the intestinal lumen. This process regulates mucus properties, can be triggered by bacterial contact, and is nonfunctional in CF due to poor mucin unfolding.protease | von Willebrand factor | gastrointestinal tract

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The substrate degradome of meprin metalloproteases reveals an unexpected proteolytic link between meprin β and ADAM10

Cited by 113 publications

References 87 publications

The anti-inflammatory peptide Ac-SDKP is released from thymosin-β4 by renal meprin-α and prolyl oligopeptidase

The anti-inflammatory peptide Ac-SDKP is released from thymosin-β4 by renal meprin-α and prolyl oligopeptidase

Metalloproteases meprin α and meprin β are C- and N-procollagen proteinases important for collagen assembly and tensile strength

Microbial-induced meprin β cleavage in MUC2 mucin and a functional CFTR channel are required to release anchored small intestinal mucus

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