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Gel LC-MS/MS analyses of MMP cleavage of tryptophanyl-tRNA synthetase

ABSTRACT: Tryptophanyl-tRNA synthetase was incubated by MMPs, cleavage products were resolved on SDS-PAGE. Cleavage products were trypsin digested and analyzed by LC-MS/MS.

INSTRUMENT(S): impact II

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Parker Jobin

LAB HEAD: Christopher Mark Overall

PROVIDER: PXD013217 | Pride | 2019-05-29

REPOSITORIES: Pride

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	180713_PJ_NS_geLCMS_w7a_D1_01_8148 (F140341).mzid_180713_PJ_NS_geLCMS_w7a_D1_01_8148_(F140341).pride.mgf.gz	Mzid
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Publications

Matrix metalloproteinases inactivate the proinflammatory functions of secreted moonlighting tryptophanyl-tRNA synthetase.

Jobin Parker G PG Solis Nestor N Machado Yoan Y Bell Peter A PA Kwon Nam Hoon NH Kim Sunghoon S Overall Christopher M CM Butler Georgina S GS

The Journal of biological chemistry 20190719 35

Tryptophanyl-tRNA synthetase (WRS) is a cytosolic aminoacyl-tRNA synthetase essential for protein synthesis. WRS is also one of a growing number of intracellular proteins that are attributed distinct noncanonical "moonlighting" functions in the extracellular milieu. Moonlighting aminoacyl-tRNA synthetases regulate processes such as inflammation, but how these multifunctional enzymes are themselves regulated remains unclear. Here, we demonstrate that WRS is secreted from human macrophages, fibrob ...[more]

PMID: 31324718

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LC-MS/MS analyses of MMP cleavage fragments of tyrosyl-tRNA synthetase

Project description:Tyrosyl-tRNA synthetase was digested with MMPs after which cleavage fragments were resolved using SDS-PAGE. Major bands were digested with trypsin and analyzed by LC-MS/MS.

2019-05-30 | PXD013197 | Pride

ATOMS analyses of MMP cleavage sites in WRS

Project description:Human tryptophanyl-tRNA synthetase (WRS) was digested by human MMPs, and digestion mixtures were subjected to terminal amine oriented mass spectrometry of substrates (ATOMS) protocol and LC-MS/MS analysis to identify MMP cleavage sites in WRS.

2019-05-29 | PXD013367 | Pride

ATOMS determination of MMP cleavage site locations in YRS

Project description:Recombinant human tyrosyl-tRNA synthetase (YRS) was cleaved using recombinant MMPs. Terminal amine oriented mass spectrometry of substrates (ATOMS) was used to identify MMP generated cleavage sites from these mixtures with LC-MS/MS analysis.

2019-05-29 | PXD013366 | Pride

Active Site Specificity Profiling of the Matrix Metalloproteinase Family: Proteomic Identification of 4,300 Cleavage Sites by MMPs 1, 2, 3, 7, 8, 9, 12, 13, and 14.

Project description:Background: The active site of matrix metalloproteinases (MMPs) is highly conserved, complicating the rational design of specific substrates and inhibitors for individual family members. Results: Active site specificity profiles of 9 MMPs were determined using high throughput Proteomic Identification of protease Cleavage Sites (PICS). Conclusion: Subtle specificity divergences distinguish individual MMP family members. Significance: Understanding individual MMP cleavage site specificities will bolster the design of MMP specific activity assays and targeted inhibitory drugs.

2015-09-29 | PXD002265 | Pride

Transcription profiling of mouse C57Bl6 MMP-7 and MMP-10 deficient mice Pseudomonas aeruginosa exposed airway epithelium.

Project description:Airway epithelium is the initial point of host-pathogen interaction in Pseudomonas aeruginosa infection, an important pathogen in cystic fibrosis and nosocomial pneumonia. We used global gene expression analysis to determine airway epithelial transcriptional responses dependent on matrilysin (MMP-7) and stromelysin-2 (MMP-10), two matrix metalloproteinases induced by acute P. aeruginosa pulmonary infection. Extraction of Differential Gene Expression (EDGE) analysis of gene expression changes in P. aeruginosa infected organotypic tracheal epithelial cell cultures from wildtype, Mmp7-/-, and Mmp10-/- mice identified 2,089 matrilysin-dependent and 1,628 stromelysin-2-dependent genes that were differentially expressed. Key node network analysis showed that these MMPs controlled distinct gene expression programs involved in proliferation, cell death, immune responses, and signal transduction, among other host defense processes. Our results demonstrate discrete roles for these MMPs in regulating epithelial responses to pseudomonas infection and show that a global genomics strategy can be used to assess MMP function. Experiment Overall Design: C57Bl6, Mmp7-/- and Mmp10-/- mouse epithelium at an organotypic air liquid interface culture was exposed to Pseudomonas aeruginosa for 1 and 24 h. RNA was collected from these samples as well as uninfected 0 h and assessed for expression changes using Affymetrix Mouse 430 2.0 Arrays. Triplicate samples were processed for each genotype at each time point.

2008-06-15 | E-GEOD-7957 | biostudies-arrayexpress

Expression data from Pseudomonas aeruginosa exposed airway epithelium from C57Bl6 and MMP-7 and MMP-10 deficient mice.

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CELF1 upregulates MMP gene expression to regulate cataract development

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2022-03-23 | GSE153022 | GEO

MiR-21 Promotes Glioma Invasion by Targeting MMP Regulators.

Project description:Substantial data indicate that microRNA-21 (miR-21) is significantly elevated in glioblastoma (GBM) and in many other tumors of various origins. This miRNA has been implicated in various aspects of carcinogenesis, including cellular proliferation, apoptosis and migration. We demonstrate that miR-21 regulates multiple genes associated with glioma cell apoptosis, migration, and invasiveness, including RECK and TIMP3, suppressors of malignancy and inhibitors of matrix metalloproteinases (MMPs). Specific inhibition of miR-21 with antisense olgonucleotides leads to elevated levels of RECK and TIMP3 and therefore reduces MMP activities in vitro and in a human model of glioma in nude mice. Moreover, down-regulation of miR-21 in glioma cells leads to decrease of their migratory and invasion abilities. Our data suggest that miR-21 contributes to the glioma malignancy by down-regulation of MMP inhibitors that leads to activation of MMPs thus promoting invasiveness of cancer cells. Our results also indicate that inhibition of a single oncomir like miR-21 with specific antisense molecules can provide a novel therapeutic approach for “physiological” modulation of multiple proteins whose expression is de-regulated in cancer.

2008-07-12 | GSE11778 | GEO

miRNA profiling in MMP-8 overexpressing MDA-MB-231 cells

Project description:Matrix metalloproteinases (MMPs) have been traditionally implicated in cancer progression because of their ability to degrade the extracellular matrix. However, some members of the MMP family have recently been identified as proteases with antitumor properties. Thus, it has been described that collagenase-2 (MMP-8) has a protective role in tumor and metastasis progression, but the molecular mechanisms underlying these effects are unknown. We show herein that Mmp8 expression causes a decrease in miR-21 levels that in turn leads to a reduction in tumor growth and lung metastasis formation by MDA-MB-231 (4175) breast cancer cells. By using both in vitro and in vivo models, we demonstrate that the mechanism responsible for these MMP-8 beneficial effects involves cleavage of decorin by MMP-8 and a subsequent reduction of transforming growth factor ? (TGF-?) signaling that controls miR-21 levels. In addition, miR-21 downregulation induced by MMP-8 increases the levels of tumor suppressors such as programmed cell death 4, which may also contribute to the decrease in tumor formation and metastasis of breast cancer cells overexpressing this metalloproteinase. These findings reveal a new signaling pathway for cancer regulation controlled by MMP-8, and contribute to clarify the molecular mechanisms by which tumor-defying proteases may exert their protective function in cancer and metastasis.

2019-01-27 | GSE124432 | GEO

Defining an invasion signature at the leading edge of cutaneous squamous cell carcinoma (SCC): IL-24 driven MMP-7 and MMP-13 expression.

Project description:Purpose: Primary cutaneous squamous cell carcinoma (SCC) can be an invasive cancer in skin and has the potential to metastasize. We aimed to define the cancer related molecular changes that distinguish non-invasive from invasive SCC. Experimental design: We used laser capture microdissection technique in combination with cDNA microarray analysis in order to determine molecular changes that associate with SCC progression. Results: We defined invasion-associated genes as those udifferentially regulated only in SCC invasive nests, but not in actinic keratosis-like dysplasia or SCC in situ regions, compared to normal epidermis. We designated these genes as “invasion signature gene set of cutaneous SCC”. Overall we found 383 up- and 354 down-regulated probe-sets that constitute the invasion signature gene set. As part of this profile, SCC invasion is associated with aberrant gene expression changes of numerous MMPs including MMP7 (FCH=5.43, FDR<0.01) and MMP13 (FCH=12.53, FDR<0.01). IL-24 is also up-regulated in the leading invasive edge of SCC (FCH=6.74, FDR<0.01). IL-24 enhanced mRNA expression of both MMP7 and MMP13 in a human SCC cell line. Laminin332, which is one of the target molecules of MMP7, had altered expression at the leading edge of SCC invasion nests at both the genomic and protein level. Conclusions: We defined the distribution of MMPs within human cutaneous SCC tissue showing distinct expression with progression from normal skin to actinic keratosis to SCC in situ to invasive carcinoma. We further suggest a potential role for IL-24 in progression to invasion via MMP7 and MMP13. Laser capture microdissection was performed on 5 cases of actinic keratosis, 5 cases of in situ SCC, and 5 cases of invasive SCC.

2013-04-30 | E-GEOD-42677 | biostudies-arrayexpress