Dataset Information

Proteomic identification of cysteine cathepsin substrates shed from the surface of cancer cells

ABSTRACT: Extracellular cysteine cathepsins are known to drive cancer progression, but besides degradation of extracellular matrix proteins little is known about their physiological substrates and thus the molecular mechanisms they deploy. One of the major mechanisms used by other extracellular proteases to facilitate cancer progression is proteolytic release of the extracellular domains of transmembrane proteins or ectodomain shedding. Here we show using a mass spectrometry-based approach that cathepsins L and S act as sheddases and cleave extracellular domains of CAM adhesion proteins and transmembrane receptors from the surface of cancer cells.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Marko Fonovic

LAB HEAD: Marko Fonovic

PROVIDER: PXD002192 | Pride | 2015-06-23

REPOSITORIES: Pride

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Publications

Proteomic Identification of Cysteine Cathepsin Substrates Shed from the Surface of Cancer Cells.

Sobotič Barbara B Vizovišek Matej M Vidmar Robert R Van Damme Petra P Gocheva Vasilena V Joyce Johanna A JA Gevaert Kris K Turk Vito V Turk Boris B Fonović Marko M

Molecular & cellular proteomics : MCP 20150616 8

Extracellular cysteine cathepsins are known to drive cancer progression, but besides degradation of extracellular matrix proteins little is known about their physiological substrates and thus the molecular mechanisms they deploy. One of the major mechanisms used by other extracellular proteases to facilitate cancer progression is proteolytic release of the extracellular domains of transmembrane proteins or ectodomain shedding. Here we show using a mass spectrometry-based approach that cathepsins ...[more]

PMID: 26081835

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Project description:The phenotypic and gene expression traits conferred by the alternative sigma factor protein M-OM-^CL in the food-borne pathogen L. monocytogenes were investigated. M-OM-^CL was shown to be important for the efficient growth of this pathogen exposed to food preservative measures such as low storage temperatures, elevated osmolarity and acidity. Based on high throughput phenotypic analysis, M-OM-^CL function was also found to be protective in L. monocytogenes EGDe cells exposed to several antimicrobial compounds including some of the antibiotics currently applied in listeriosis treatment. The expression of flagella genes and motility were significantly compromized upon loss of M-OM-^CL function. A comparative transcriptome analysis of exponential growth EGDe wild type and sigL null cells unveiled 394 genes that are positively controlled through M-OM-^CL dependent transcriptional regulation mechanisms in this bacterium during growth at low (3M-BM-0C) and optimized (37M-BM-0C) temperature conditions. Genes identified indicate that M-OM-^CL is a pleitropic transcription regulator mediating positive expression control of genes involved in diverse cellular processes including protein synthesis, molecular transport, energy metabolism, respiration, transcription regulation, metabolite biosynthesis, and cell envelope composition modification. Overall our observations have revealed that the loss of M-OM-^CL function leads to extensive gene expression defects in L. monocytogenes EGDe cells, and these are consistent with compromized nutrient assimilation, energy metabolism, protein synthesis, and metabolite biosynthesis processes as well as altered cell envelope composition and motility. Numerous gene expression changes imposed by M-OM-^CL loss in EGDe are thus also consistent with pleiotropic phenotypic defects detected in the L. monocytogenes EGDe M-bM-^HM-^FsigL strain. Gene expression DNA-microarray. Two-color hybridizations on 8*15K Agilent arrays. Wild type and three KO mutants in three temperatures, three biological replicates in each condition.

Project description:The two-component system CBO0366/CBO0365 was recently demonstrated to have a role in cold tolerance of Group I Clostridium botulinum ATCC 3502. The mechanisms under its control, ultimately resulting in increased sensitivity to low temperature, are unknown. A transcriptomic analysis with DNA microarrays was performed to identify the differences in global gene expression patterns of the wild-type ATCC 3502 and a derivative mutant with insertionally inactivated cbo0365 at 37 M-BM-0C and 15 M-BM-0C. Altogether 150 or 141 chromosomal CDSs were found to be differently expressed in the cbo0365 mutant at 37 M-BM-0C or 15 M-BM-0C, respectively, and thus considered to be under direct or indirect transcriptional control of the response regulator CBO0365. Of the differentially-expressed CDSs, expression of 141 CDSs was similarly affected at both temperatures investigated, suggesting that the putative CBO0365 regulon was practically not affected by temperature. The regulon involved genes related to acetone-butanol-ethanol (ABE) fermentation, motility, to arsenic resistance, and phosphate uptake and transport. Deteriorated growth at 17 M-BM-0C was observed for mutants with disrupted ABE fermentation pathway components (crt, bdh and ctfA), arsenic detoxifying machinery components (arsC and arsR), or phosphate uptake mechanism components (phoT), suggesting roles for these mechanisms in cold tolerance of Group I C. botulinum. Electrophoretic mobility shift assays showed recombinant CBO0365 to bind to the promoter regions of crt, arsR, and phoT, as well as to the promoter region of its own operon, suggesting direct DNA-binding transcriptional activation or repression as means for CBO0365 in regulating these operons. The results provide insight to the mechanisms Group I C. botulinum utilize in coping with cold. C. botulinum ATCC 3502 cbo0365 mutant vs. wild type; 3 replicates of each strain; growth at 37C in TPGY broth batch culture and subjected to cold shock to 15C; sampling at mid-log growth phase before cold shock, and 1 h after temperature downshift to 15C (= 2 time points). Dye-swapped hybridization.

Dataset Information

Proteomic identification of cysteine cathepsin substrates shed from the surface of cancer cells

Publications

Proteomic Identification of Cysteine Cathepsin Substrates Shed from the Surface of Cancer Cells.

Similar Datasets

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