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Cells Dec 2019The extracellular matrix (ECM) is a complex and specialized three-dimensional macromolecular network, present in nearly all tissues, that also interacts with cell... (Review)
Review
The extracellular matrix (ECM) is a complex and specialized three-dimensional macromolecular network, present in nearly all tissues, that also interacts with cell surface receptors on joint resident cells. Changes in the composition and physical properties of the ECM lead to the development of many diseases, including osteoarthritis (OA). OA is a chronic degenerative rheumatic disease characterized by a progressive loss of synovial joint function as a consequence of the degradation of articular cartilage, also associated with alterations in the synovial membrane and subchondral bone. During OA, ECM-degrading enzymes, including urokinase-type plasminogen activator (uPA), matrix metalloproteinases (MMPs), and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs), cleave ECM components, such as fibronectin (Fn), generating fibronectin fragments (Fn-fs) with catabolic properties. In turn, Fn-fs promote activation of these proteinases, establishing a degradative and inflammatory feedback loop. Thus, the aim of this review is to update the contribution of ECM-degrading proteinases to the physiopathology of OA as well as their modulation by Fn-fs.
Topics: ADAMTS Proteins; Animals; Cartilage, Articular; Endopeptidases; Extracellular Matrix; Fibronectins; Humans; Matrix Metalloproteinases; Metalloendopeptidases; Osteoarthritis; Peptide Hydrolases
PubMed: 31877874
DOI: 10.3390/cells9010040 -
British Journal of Cancer Jun 2023Prostate cancer is the most common cancer in men in the developed world, with most deaths caused by advanced and metastatic disease which has no curative options. Here,...
BACKGROUND
Prostate cancer is the most common cancer in men in the developed world, with most deaths caused by advanced and metastatic disease which has no curative options. Here, we identified Mbtps2 alteration to be associated with metastatic disease in an unbiased in vivo screen and demonstrated its regulation of fatty acid and cholesterol metabolism.
METHODS
The Sleeping Beauty transposon system was used to randomly alter gene expression in the Pten murine prostate. MBTPS2 was knocked down by siRNA in LNCaP, DU145 and PC3 cell lines, which were then phenotypically investigated. RNA-Seq was performed on LNCaP cells lacking MBTPS2, and pathways validated by qPCR. Cholesterol metabolism was investigated by Filipin III staining.
RESULTS
Mbtps2 was identified in our transposon-mediated in vivo screen to be associated with metastatic prostate cancer. Silencing of MBTPS2 expression in LNCaP, DU145 and PC3 human prostate cancer cells reduced proliferation and colony forming growth in vitro. Knockdown of MBTPS2 expression in LNCaP cells impaired cholesterol synthesis and uptake along with reduced expression of key regulators of fatty acid synthesis, namely FASN and ACACA.
CONCLUSION
MBTPS2 is implicated in progressive prostate cancer and may mechanistically involve its effects on fatty acid and cholesterol metabolism.
Topics: Male; Humans; Animals; Mice; Lipogenesis; Sterol Regulatory Element Binding Protein 1; Cell Line, Tumor; Prostatic Neoplasms; Cholesterol; Fatty Acids; Cell Proliferation; Gene Expression Regulation, Neoplastic; Metalloendopeptidases
PubMed: 36991255
DOI: 10.1038/s41416-023-02237-7 -
Biochimica Et Biophysica Acta.... Jan 2022The metalloproteinase meprin β plays an important role during collagen I deposition in the skin, mucus detachment in the small intestine and also regulates the... (Review)
Review
The metalloproteinase meprin β plays an important role during collagen I deposition in the skin, mucus detachment in the small intestine and also regulates the abundance of different cell surface proteins such as the interleukin-6 receptor (IL-6R), the triggering receptor expressed on myeloid cells 2 (TREM2), the cluster of differentiation 99 (CD99), the amyloid precursor protein (APP) and the cluster of differentiation 109 (CD109). With that, regulatory mechanisms that control meprin β activity and regulate its release from the cell surface to enable access to distant substrates are increasingly important. Here, we will summarize factors that alternate meprin β activity and thereby regulate its proteolytic activity on the cell surface or in the supernatant. We will also discuss cleavage of the IL-6R and TREM2 on the cell surface and compare it to CD109. CD109, as a substrate of meprin β, is cleaved within the protein core, thereby releasing defined fragments from the cell surface. At last, we will also summarize the role of proteases in general and meprin β in particular in substrate release on extracellular vesicles.
Topics: Animals; Extracellular Vesicles; Humans; Metalloendopeptidases; Proteolysis; Signal Transduction
PubMed: 34626678
DOI: 10.1016/j.bbamcr.2021.119136 -
Molecules (Basel, Switzerland) Feb 2023Ochratoxin A (OTA) is considered one of the main mycotoxins responsible for health problems and considerable economic losses in the feed industry. The aim was to study...
Ochratoxin A (OTA) is considered one of the main mycotoxins responsible for health problems and considerable economic losses in the feed industry. The aim was to study OTA's detoxifying potential of commercial protease enzymes: (i) bromelain cysteine-protease, (ii) bovine trypsin serine-protease and (iii) neutral metalloendopeptidase. In silico studies were performed with reference ligands and T-2 toxin as control, and in vitro experiments. In silico study results showed that tested toxins interacted near the catalytic triad, similar to how the reference ligands behave in all tested proteases. Likewise, based on the proximity of the amino acids in the most stable poses, the chemical reaction mechanisms for the transformation of OTA were proposed. In vitro experiments showed that while bromelain reduced OTA's concentration in 7.64% at pH 4.6; trypsin at 10.69% and the neutral metalloendopeptidase in 8.2%, 14.44%, 45.26% at pH 4.6, 5 and 7, respectively ( < 0.05). The less harmful α-ochratoxin was confirmed with trypsin and the metalloendopeptidase. This study is the first attempt to demonstrate that: (i) bromelain and trypsin can hydrolyse OTA in acidic pH conditions with low efficiency and (ii) the metalloendopeptidase was an effective OTA bio-detoxifier. This study confirmed α-ochratoxin as a final product of the enzymatic reactions in real-time practical information on OTA degradation rate, since in vitro experiments simulated the time that food spends in poultry intestines, as well as their natural pH and temperature conditions.
Topics: Animals; Cattle; Ochratoxins; Bromelains; Molecular Docking Simulation; Trypsin; Mycotoxins; Animal Feed; Metalloendopeptidases
PubMed: 36903263
DOI: 10.3390/molecules28052019 -
Clinical Dysmorphology Apr 2023Restrictive dermopathy (RD) (OMIM 275210) is a rare, lethal genodermatosis belonging to the group of laminopathies. It is caused by biallelic variants in , which is...
Restrictive dermopathy (RD) (OMIM 275210) is a rare, lethal genodermatosis belonging to the group of laminopathies. It is caused by biallelic variants in , which is involved in lamin A post-translational processing or, less frequently, by monoallelic variants in , leading to accumulation of truncated prelamin A protein (Navarro et al., 2004; Navarro et al., 2005). The main characteristics of RD include intrauterine growth retardation (IUGR), reduced fetal movement, premature rupture of membranes, translucent rigid skin, dysmorphic features and joint contractures. The prognosis is poor with all reported cases resulting in stillbirth or neonatal death (Navarro et al., 2014). Herein we report a neonate born to healthy, non-consanguineous parents from Greece. The pregnancy was uneventful until the 32 week, when a routine scan showed severe fetal growth restriction with normal Doppler flows. The female proband was born at 33 weeks of gestation by caesarean section, due to premature rupture of membranes, as well as anhydramnios, IUGR, fetal hypokinesia and distress. Her birth weight was 1.36 kg (5 centile, −1.6SD), length was 41 cm (14 centile) and head circumference was 29 cm (14 centile). Apgar score was 4 and 8 at the 1 and 5 minutes, respectively. She required immediate intubation and admission to the neonatal intensive care unit. She had a large fontanelle, short palpebral fissures, a small pinched nose, low-set dysplastic ears and an open, O-shaped mouth (Fig. 1). She had multiple joint contractures. Her skin was rigid and translucent and progressively developed erosions and scaling. She did not have eyebrows or eyelashes. She had severe lung hypoplasia and died of respiratory insufficiency on the 22 day of life.
Topics: Humans; Membrane Proteins; Metalloendopeptidases
PubMed: 36876346
DOI: 10.1097/MCD.0000000000000453 -
The Plant Journal : For Cell and... Apr 2021Protein homeostasis (proteostasis) is crucial for proper cellular function, including the production of peptides with biological functions through controlled...
Protein homeostasis (proteostasis) is crucial for proper cellular function, including the production of peptides with biological functions through controlled proteolysis. Proteostasis has roles in maintenance of cellular functions and plant interactions with the environment under physiological conditions. Plant stress continues to reduce agricultural yields causing substantial economic losses; thus, it is critical to understand how plants perceive stress signals to elicit responses for survival. As previously shown in Arabidopsis thaliana, thimet oligopeptidases (TOPs) TOP1 (also referred to as organellar oligopeptidase) and TOP2 (also referred to as cytosolic oligopeptidase) are essential components in plant response to pathogens, but further characterization of TOPs and their peptide substrates is required to understand their contributions to stress perception and defense signaling. Herein, label-free peptidomics via liquid chromatography-tandem mass spectrometry was used to differentially quantify 1111 peptides, originating from 369 proteins, between the Arabidopsis Col-0 wild type and top1top2 knock-out mutant. This revealed 350 peptides as significantly more abundant in the mutant, representing accumulation of these potential TOP substrates. Ten direct substrates were validated using in vitro enzyme assays with recombinant TOPs and synthetic candidate peptides. These TOP substrates are derived from proteins involved in photosynthesis, glycolysis, protein folding, biogenesis, and antioxidant defense, implicating TOP involvement in processes aside from defense signaling. Sequence motif analysis revealed TOP cleavage preference for non-polar residues in the positions surrounding the cleavage site. Identification of these substrates provides a framework for TOP signaling networks, through which the interplay between proteolytic pathways and defense signaling can be further characterized.
Topics: Arabidopsis; Arabidopsis Proteins; Metalloendopeptidases; Proteolysis
PubMed: 33481299
DOI: 10.1111/tpj.15165 -
Microbiology and Immunology Jan 2017Vibrio vulnificus, a gram-negative halophilic estuarine bacterium, is an opportunistic human pathogen that causes rapidly progressive fatal septicemia and necrotizing... (Review)
Review
Vibrio vulnificus, a gram-negative halophilic estuarine bacterium, is an opportunistic human pathogen that causes rapidly progressive fatal septicemia and necrotizing wound infection. This species also causes hemorrhagic septicemia called vibriosis in cultured eels. It has been proposed that a range of virulence factors play roles in pathogenesis during human and/or eel infection. Among these factors, a metalloprotease (V. vulnificus protease [VVP]) and a cytolytic toxin (V. vulnificus hemolysin [VVH]) are of significant importance. VVP elicits the characteristic edematous and hemorrhagic skin damage, whereas VVH exhibits powerful hemolytic and cytolytic activities and contributes to bacterial invasion from the intestine to the blood stream. In addition, a few V. vulnificus strains isolated from diseased eels have recently been found to produce a serine protease designated as V. vulnificus serine protease (VvsA) instead of VVP. Similarly to VVP, VvsA may possess various toxic activities such as collagenolytic, cytotoxic and edema-forming activity. In this review, regulation of V. vulnificus VVP, VVH and VvsA is clarified in terms of expression at the mRNA and protein levels. The explanation is given on the basis of the quorum sensing system, which is dependent on bacterial cell density. In addition, the roles of environmental factors and global regulators, such as histone-like nucleoid structuring protein, cyclic adeno monophosphate receptor protein, RpoS, HlyU, Fur, ToxRS, AphB and LeuO, in this regulation are outlined. The cumulative impact of these regulatory systems on the pathogenicity of V. vulnificus is here delineated.
Topics: Animals; Bacterial Proteins; Hemolysin Proteins; Humans; Metalloendopeptidases; Metalloproteases; RNA, Messenger; Vibrio vulnificus; Virulence Factors
PubMed: 28111826
DOI: 10.1111/1348-0421.12465 -
ELife Mar 2021Endoplasmic reticulum (ER) and mitochondria form close physical associations to facilitate calcium transfer, thereby regulating mitochondrial function. Neurons with high...
Endoplasmic reticulum (ER) and mitochondria form close physical associations to facilitate calcium transfer, thereby regulating mitochondrial function. Neurons with high metabolic demands, such as sensory hair cells, are especially dependent on precisely regulated ER-mitochondria associations. We previously showed that the secreted metalloprotease pregnancy-associated plasma protein-aa (Pappaa) regulates mitochondrial function in zebrafish lateral line hair cells (Alassaf et al., 2019). Here, we show that mutant hair cells exhibit excessive and abnormally close ER-mitochondria associations, suggesting increased ER-mitochondria calcium transfer. mutant hair cells are more vulnerable to pharmacological induction of ER-calcium transfer. Additionally, mutant hair cells display ER stress and dysfunctional downstream processes of the ER-mitochondria axis including altered mitochondrial morphology and reduced autophagy. We further show that Pappaa influences ER-calcium transfer and autophagy via its ability to stimulate insulin-like growth factor-1 bioavailability. Together our results identify Pappaa as a novel regulator of the ER-mitochondria axis.
Topics: Animals; Calcium; Endoplasmic Reticulum; Endoplasmic Reticulum Stress; Hair Cells, Auditory, Inner; Lateral Line System; Metalloendopeptidases; Microscopy, Electron, Scanning Transmission; Mitochondria; Zebrafish; Zebrafish Proteins
PubMed: 33759764
DOI: 10.7554/eLife.59687 -
Matrix Biology : Journal of the... 2015Metalloproteases meprin α and meprin β were recently discovered as procollagen proteinases, capable of cleaving off the globular C- and N-terminal prodomains of... (Review)
Review
Metalloproteases meprin α and meprin β were recently discovered as procollagen proteinases, capable of cleaving off the globular C- and N-terminal prodomains of fibrillar collagen type I and type III. This proteolytic process is indeed sufficient to induce collagen fibril assembly as visualized by transmission electron microscopy. The biological relevance was demonstrated with the help of meprin α and meprin β knock-out mice, which exhibit decreased collagen deposition in skin resulting in impaired tensile strength. On the other hand, overexpression of meprin metalloproteases was found under fibrotic conditions in the skin (keloids) and the lung (pulmonary hypertension). Thus, regulation of meprin activity by specific inhibition to reduce collagen maturation might be a suitable approach for the treatment of certain pathological conditions.
Topics: Animals; Collagen Type I; Collagen Type III; Gene Expression Regulation, Enzymologic; Gene Knockout Techniques; Humans; Hypertension, Pulmonary; Keloid; Metalloendopeptidases; Mice; Procollagen; Tensile Strength
PubMed: 25617491
DOI: 10.1016/j.matbio.2015.01.010 -
Journal of Translational Medicine Mar 2021The MBTPS2 gene on the X-chromosome encodes the membrane-bound transcription factor protease, site-2 (MBTPS2) or site-2 protease (S2P) which cleaves and activates... (Review)
Review
The MBTPS2 gene on the X-chromosome encodes the membrane-bound transcription factor protease, site-2 (MBTPS2) or site-2 protease (S2P) which cleaves and activates several signaling and regulatory proteins from the membrane. The MBTPS2 is critical for a myriad of cellular processes, ranging from the regulation of cholesterol homeostasis to unfolded protein responses. While its functional role has become much clearer in the recent years, how mutations in the MBTPS2 gene lead to several human disorders with different phenotypes including Ichthyosis Follicularis, Atrichia and Photophobia syndrome (IFAP) with or without BRESHECK syndrome, Keratosis Follicularis Spinulosa Decalvans (KFSD), Olmsted syndrome, and Osteogenesis Imperfecta type XIX remains obscure. This review presents the biological role of MBTPS2 in development, summarizes its mutations and implicated disorders, and discusses outstanding unanswered questions.
Topics: Humans; Metalloendopeptidases; Mutation, Missense; Pedigree; Peptide Hydrolases; Transcription Factors
PubMed: 33743732
DOI: 10.1186/s12967-021-02779-5