scholarly journals The Heat Shock Protein 60 and Pap1 Participate in the Sporothrix schenckii-Host Interaction

2021 ◽  
Vol 7 (11) ◽  
pp. 960
Author(s):  
Laura C. García-Carnero ◽  
Roberta Salinas-Marín ◽  
Nancy E. Lozoya-Pérez ◽  
Katarzyna Wrobel ◽  
Kazimierz Wrobel ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Wall ◽  
Sporothrix Schenckii ◽  
Type I ◽  
Heat Shock Protein 60 ◽  
Uncharacterized Protein ◽  
Adhesive Properties ◽  
Lethal Challenge ◽  
Host Interaction ◽  
Extracellular Matrix Components

Sporothrixschenckii is one of the etiological agents of sporotrichosis, a worldwide-distributed subcutaneous mycosis. Its cell wall contains a glycoconjugate composed of rhamnose, mannose, glucuronic acid, and proteins, named peptidorhamnomannan, which harbors important Sporothrix-specific immunogenic epitopes. Although the peptidorhamnomannan carbohydrate moiety has been extensively studied, thus far, little is known about the protein core. Here, using LC-MS/MS, we analyzed the S.schenckii peptidorhamnomannan peptide fraction and generated mass signals of 325 proteins, most of them likely to be moonlighting proteins. Among the identified proteins, chaperonin GroEL/Hsp60 and the uncharacterized protein Pap1 were selected for further analysis. Both proteins were heterologously expressed in bacteria, and they showed adhesive properties to the extracellular matrix proteins laminin, elastin, fibrinogen, and fibronectin, although Pap1 also was bound to type-I and type-II collagen. The inoculation of concentrations higher than 40 μg of these proteins, separately, increased immune effectors in the hemolymph of Galleriamellonella larvae and protected animals from an S.schenckii lethal challenge. These observations were confirmed when yeast-like cells, pre-incubated with anti-rHsp60 or anti-rPap1 antibodies were used to inoculate larvae. The animals inoculated with pretreated cells showed increased survival rates when compared to the control groups. In conclusion, we report that Hsp60 and Pap1 are part of the cell wall peptidorhamnomannan, can bind extracellular matrix components, and contribute to the S.schenckii virulence. To our knowledge, this is the first report about moonlighting protein in the S.schenckii cell wall with an important role during the pathogen–host interaction.

scholarly journals Glyceraldehyde-3-Phosphate Dehydrogenase of Paracoccidioides brasiliensis Is a Cell Surface Protein Involved in Fungal Adhesion to Extracellular Matrix Proteins and Interaction with Cells

2006 ◽  
Vol 74 (1) ◽  
pp. 382-389 ◽  
Author(s):  
Mônica Santiago Barbosa ◽  
Sônia Nair Báo ◽  
Patrícia Ferrari Andreotti ◽  
Fabrícia P. de Faria ◽  
Maria Sueli S. Felipe ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Wall ◽  
Western Blot ◽  
Polyclonal Antibody ◽  
Type I Collagen ◽  
Paracoccidioides Brasiliensis ◽  
Host Cells ◽  
Yeast Cells ◽  
Type I ◽  
Extracellular Matrix Components

ABSTRACT The pathogenic fungus Paracoccidioides brasiliensis causes paracoccidioidomycosis, a pulmonary mycosis acquired by inhalation of fungal airborne propagules, which may disseminate to several organs and tissues, leading to a severe form of the disease. Adhesion to and invasion of host cells are essential steps involved in the infection and dissemination of pathogens. Furthermore, pathogens use their surface molecules to bind to host extracellular matrix components to establish infection. Here, we report the characterization of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of P. brasiliensis as an adhesin, which can be related to fungus adhesion and invasion. The P. brasiliensis GAPDH was overexpressed in Escherichia coli, and polyclonal antibody against this protein was obtained. By immunoelectron microscopy and Western blot analysis, GAPDH was detected in the cytoplasm and the cell wall of the yeast phase of P. brasiliensis. The recombinant GAPDH was found to bind to fibronectin, laminin, and type I collagen in ligand far-Western blot assays. Of special note, the treatment of P. brasiliensis yeast cells with anti-GAPDH polyclonal antibody and the incubation of pneumocytes with the recombinant protein promoted inhibition of adherence and internalization of P. brasiliensis to those in vitro-cultured cells. These observations indicate that the cell wall-associated form of the GAPDH in P. brasiliensis could be involved in mediating binding of fungal cells to fibronectin, type I collagen, and laminin, thus contributing to the adhesion of the microorganism to host tissues and to the dissemination of infection.

scholarly journals Lactobacillus Cell Surface Proteins Involved in Interaction with Mucus and Extracellular Matrix Components

2020 ◽  
Vol 77 (12) ◽  
pp. 3831-3841
Author(s):  
Lidia Muscariello ◽  
Barbara De Siena ◽  
Rosangela Marasco
Keyword(s):  
Extracellular Matrix ◽  
Cell Surface ◽  
Intestinal Flora ◽  
Surface Proteins ◽  
Matrix Proteins ◽  
Cell Surface Proteins ◽  
Host Interaction ◽  
Extracellular Matrix Components ◽  
Probiotic Strains ◽  
Matrix Components

AbstractThe gut microbiota is a complex microbial ecosystem where bacteria, through mutual interactions, cooperate in maintaining of wellbeing and health. Lactobacilli are among the most important constituents of human and animal intestinal microbiota and include many probiotic strains. Their presence ensures protection from invasion of pathogens, as well as stimulation of the immune system and protection of the intestinal flora, often exerted through the ability to interact with mucus and extracellular matrix components. The main factors responsible for mediating adhesion of pathogens and commensals to the gut are cell surface proteins that recognize host targets, as mucus layer and extracellular matrix proteins. In the last years, several adhesins have been reported to be involved in lactobacilli–host interaction often miming the same mechanism used by pathogens.

scholarly journals TEF-1 and C/EBPβ are major p38α MAPK-regulated transcription factors in proliferating cardiomyocytes

2006 ◽  
Vol 396 (1) ◽  
pp. 163-172 ◽  
Author(s):  
Concetta Ambrosino ◽  
Tomoko Iwata ◽  
Claudio Scafoglio ◽  
Massimo Mallardo ◽  
Rüdiger Klein ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Transcription Factors ◽  
Environmental Stress ◽  
Type I Collagen ◽  
Specific Gene ◽  
Type I ◽  
Transcriptional Enhancer ◽  
Extracellular Matrix Components ◽  
Mitogen Activated Protein ◽  
P38 Mapks

p38 MAPKs (mitogen-activated protein kinases) play important roles in the regulation of cellular responses to environmental stress. Recently, this signalling pathway has also been implicated in the regulation of processes unrelated to stress, for example, in T lymphocytes and cardiomyocytes. In order to identify molecular targets responsible for the housekeeping functions of p38 MAPKs, we have analysed the differences in the transcriptomes of normally proliferating wild-type and p38α knockout immortalized embryonic cardiomyocytes. Interestingly, many potential components of the myocardium extracellular matrix were found to be upregulated in the absence of p38α. Further analysis of the microarray data identified TEF-1 (transcriptional enhancer factor-1), a known regulator of heart-specific gene expression, and C/EBPβ (CCAAT/enhancer-binding protein β), as the two transcription factors the binding sites of which were most enriched in the promoters of p38α-regulated genes. We have focused on the study of the extracellular matrix component COL1A1 (α1 chain of type I collagen) and found evidence for the involvement of both TEF-1 and C/EBPβ in the p38α-dependent inhibition of COL1A1 transcription. Our data therefore show that p38 MAPKs regulate TEF-1 and C/EBPβ transcriptional activity in the absence of environmental stress and suggests a role for p38α in the expression of extracellular matrix components that maintain organ architecture.

scholarly journals Expression of extracellular matrix components is regulated by substratum.

1990 ◽  
Vol 110 (4) ◽  
pp. 1405-1415 ◽  
Author(s):  
C H Streuli ◽  
M J Bissell
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Type I Collagen ◽  
Basement Membranes ◽  
Type I ◽  
Collagen Gels ◽  
Extracellular Matrix Components ◽  
Matrix Components ◽  
Cells Cultured

Reconstituted basement membranes and extracellular matrices have been demonstrated to affect, positively and dramatically, the production of milk proteins in cultured mammary epithelial cells. Here we show that both the expression and the deposition of extracellular matrix components themselves are regulated by substratum. The steady-state levels of the laminin, type IV collagen, and fibronectin mRNAs in mammary epithelial cells cultured on plastic dishes and on type I collagen gels have been examined, as has the ability of these cells to synthesize, secrete, and deposit laminin and other, extracellular matrix proteins. We demonstrate de novo synthesis of a basement membrane by cells cultured on type I collagen gels which have been floated into the medium. Expression of the mRNA and proteins of basement membranes, however, are quite low in these cultures. In contrast, the levels of laminin, type IV collagen, and fibronectin mRNAs are highest in cells cultured on plastic surfaces, where no basement membrane is deposited. It is suggested that the interaction between epithelial cells and both basement membrane and stromally derived matrices exerts a negative influence on the expression of mRNA for extracellular matrix components. In addition, we show that the capacity for lactational differentiation correlates with conditions that favor the deposition of a continuous basement membrane, and argue that the interaction between specialized epithelial cells and stroma enables them to create their own microenvironment for accurate signal transduction and phenotypic function.

scholarly journals Morphometric analysis of the human endoneurial extracellular matrix components during aging

2021 ◽  
Vol 73 (1) ◽  
pp. 103-110
Author(s):  
Braca Kundalic ◽  
Sladjana Ugrenovic ◽  
Ivan Jovanovic ◽  
Vladimir Petrovic ◽  
Aleksandar Petrovic ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Collagen Type ◽  
Linear Regression Analysis ◽  
Collagen Type I ◽  
Nerve Fibers ◽  
Collagen Type Iv ◽  
Type I ◽  
Type Iv ◽  
Ecm Proteins ◽  
Extracellular Matrix Components

The aim of this study was to analyze the expression of extracellular matrix (ECM) proteins in human endoneurium during aging. We harvested 15 cadaveric sural nerves, distributed in 3 age groups (I: 25-44, II: 45-64, III: 65-86 years old). Histological sections were stained immunohistochemically for the presence of collagen type I, type IV and laminin, and the ImageJ processing program was used in morphometrical analysis to determine the percentages of these endoneurial proteins. In two younger groups, the endoneurial matrix of the sural nerve was composed from about equal proportions of these proteins, which may be considered a favorable microenvironment for the regeneration of nerve fibers. Linear regression analysis showed a significant increase in endoneurial collagen type IV with age, while collagen type I and laminin significantly decreased during the aging process. In cases older than 65 years, remodeling of the endoneurial matrix was observed to be significantly higher for the presence of collagen type IV, and lower for the expression of collagen type I and laminin. This age-related imbalance of ECM proteins could represent a disadvantageous microenvironment for nerve fiber regeneration in older adults. Our findings contribute to the development of therapeutic approaches for peripheral nerve regeneration.

scholarly journals Presence of ROS in Inflammatory Environment of Peri-Implantitis Tissue: In Vitro and In Vivo Human Evidence

2019 ◽  
Vol 9 (1) ◽  
pp. 38
Author(s):  
Eitan Mijiritsky ◽  
Letizia Ferroni ◽  
Chiara Gardin ◽  
Oren Peleg ◽  
Alper Gultekin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Mesenchymal Stem Cells ◽  
Human Model ◽  
Inflammatory Cell Infiltrate ◽  
Mitochondrial Activity ◽  
Type I ◽  
Biological Organization ◽  
Extracellular Matrix Components ◽  
Matrix Components

Analyses of composition, distribution of cellular and extracellular matrix components, and molecular analysis of mitochondria related genes of bone loss in the presence of inflammatory environment in humans was the aim of the present project. As a human model we chose peri-implantitis. Morphological analyses were performed by means classical histological, immunohistochemical, and SEM (scanning electron miscroscopy) test. Gene expression analysis was performed to evaluate epithelium maturation, collagen fiber production, and genes related to mitochondrial activity. It was found that a well-defined keratinocyte epithelium was present on the top of all specimens; a distinct basal lamina was present, as well as desmosomes and autophagic processes related to the maturation of keratinocytes. Under this epithelium, a full inflammatory cell infiltrate was present for about 60% of the represented by plasma cells. Collagen type I fibers were present mainly in the form of fragmented cord tissue without cells. A different distribution of blood vessels was also present from the apical to the most coronal portion of the specimens. High levels of genes related to oxidative stress were present, as well as the activation of genes related to the loss of ability of osteogenic commitment of Mesenchymal stem cells into osteoblasts. Our study suggests that peri-implantitis lesions exhibit a well defined biological organization not only in terms of inflammatory cells but also on vessel and extracellular matrix components even if no difference in the epithelium is evident, and that the presence of reactive oxygen species (ROS) related to the inflammatory environment influences the correct commitment of Mesenchymal stem cells.

scholarly journals A Trypanosoma cruzi-secreted 80 kDa proteinase with specificity for human collagen types I and IV

1997 ◽  
Vol 325 (1) ◽  
pp. 129-137 ◽  
Author(s):  
Jaime Martins SANTANA ◽  
Philippe GRELLIER ◽  
Joseph SCHRÉVEL ◽  
Antonio R. L. TEIXEIRA
Keyword(s):  
Extracellular Matrix ◽  
Enzyme Activity ◽  
Chagas Disease ◽  
Mammalian Cells ◽  
Peptide Sequence ◽  
Proteinase Activity ◽  
Type I ◽  
Collagen Types ◽  
Small Proteins ◽  
Extracellular Matrix Components

Specific interactions between parasites and extracellular matrix components are an important mechanism in the dissemination of Chagas' disease. Binding of the extracellular matrix proteins to Trypanosoma cruzireceptors has been described as a significant step in this phenomenon. In this study, a specific proteinase activity was identified in cell-free extracts of amastigote, trypomastigote and epimastigote forms of T. cruziusing the collagenase fluorogenic substrate N-Suc-Gly-Pro-Leu-Gly-Pro-7-amido-4-methylcoumarin. Isolation of this activity was achieved by a four-step FPLC procedure. Optimal enzyme activity was found to occur at pH 8.0 and was associated with a single T. cruzi80 kDa protein (Tc 80 proteinase) on SDS/PAGE under reducing conditions. An internal peptide sequence of Tc 80 proteinase was obtained (AGDNYTPPE), and no similarity was found to previously described proteinases of T. cruzi. This enzyme activity is strongly inhibited by HgCl2, tosyl-lysylchloromethane (‘TLCK’) p-chloromercuribenzoate and benzyloxycarbonyl-Phe-Ala-diazomethane. The purified enzyme was able to hydrolyse purified human [14C]collagen types I and IV at neutral pH, but not 14C-labelled BSA, rat laminin, rabbit IgG or small proteins such as insulin or cytochrome c. In addition, Tc 80 proteinase activity was found to be secreted by T. cruziforms infective to mammalian cells. Furthermore we demonstrated that purified Tc 80 proteinase mediates native collagen type I hydrolysis in rat mesentery. This feature is compared with that of Clostridium histolyticum collagenase. These findings suggest that Tc 80 proteinase may facilitate T. cruzihost-cell infection by degrading the collagens of the extracellular matrix and could represent a good target for Chagas' disease chemotherapy.

scholarly journals Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix production.

1982 ◽  
Vol 92 (2) ◽  
pp. 452-461 ◽  
Author(s):  
J E Aubin ◽  
J N Heersche ◽  
M J Merrilees ◽  
J Sodek
Keyword(s):  
Extracellular Matrix ◽  
Bone Cells ◽  
Bone Cell ◽  
Population Doubling ◽  
Cell Populations ◽  
Type I ◽  
Mixed Cell ◽  
Extracellular Matrix Components ◽  
Wide Range

Clones of nontransformed hormone-responsive bone cells have been isolated in vitro from mixed cell populations of fetal rat calvaria. In several independent isolations, microscopically visible colonies appeared at plating efficiencies of 5-10% of the starting cell numbers. Of these clones, approximately 10% grew to mass populations which could be assayed for a number of growth and biochemical properties. Although some similarities existed among the clones, they could be distinguished from each other and from the mixed cell populations. Population-doubling times (tDs) and saturation densities varied over a wide range: e.g., tDs of 24-72 h and saturation densities of 0.4-5 x 10(5) cells/cm2. Morphologies varied from roughly polygonal multilayering cells to typically spindle-shaped monolayering cells. Hormone responsiveness, as measured by stimulation of cAMP by hormones, indicated that some clones were responsive to both parathyroid hormone (PTH) and prostaglandin E2 (PGE2), while others responded to PTH only. Analysis of extracellular matrix components revealed that all clones produced type I and type III collagens, though in different proportions. Similarly, although all clones synthesized four glycosaminoglycans (hyaluronic acid, heparan sulfate, chondroitin sulfate, and dermatan sulfate), the quantities of each were distinctive from clone to clone. Further investigation of such clones is continuing to define more precisely the heterogeneity of clonal bone cell populations in vitro. They represent an important step in the study of the endocrinology and differentiation of bone.

Effect of AT2 blockade on cardiac hypertrophy as induced by high dietary salt in the proatrial natriuretic peptide (ANP) gene-disrupted mouse

2006 ◽  
Vol 84 (6) ◽  
pp. 625-634 ◽  
Author(s):  
Ekaterini Angelis ◽  
M. Yat Tse ◽  
Michael A. Adams ◽  
Stephen C. Pang
Keyword(s):  
Extracellular Matrix ◽  
Cardiac Hypertrophy ◽  
Natriuretic Peptide ◽  
Transforming Growth Factor ◽  
Matrix Metalloproteinase 2 ◽  
Marker Genes ◽  
Type I ◽  
Dietary Salt ◽  
Tissue Mass ◽  
Extracellular Matrix Components

The role of the angiotensin II type 2 receptor (AT2) during alterations in cardiac size remains largely unclear. Through employment of an AT2 antagonist, the present study explored a possible involvement of the AT2 receptor during salt-induced cardiac hypertrophy in the proatrial natriuretic peptide gene-disrupted mouse (ANP−/−). ANP−/− mice received either saline solution or the AT2 antagonist, PD123319, and were then placed on a high salt diet (8.0% NaCl) for 3 weeks. Cardiac and pulmonary size, expression of the renin–angiotensin system (RAS), and the behaviour of various hypertrophy marker genes were assessed. PD123319 caused enhanced expression of the systemic RAS, yet the cardiac RAS was largely unaffected. Although AT2 blockade did not alter whole cardiac mass, right ventricle mass, as well as pulmonary mass-to-body mass ratios were significantly decreased. Collagen type I was decreased in the latter tissues, likely contributing to the regression in mass. Several players essential in the maintenance of myocardial extracellular matrix homeostasis including B-type natriuretic peptide, matrix metalloproteinase-2, tumour necrosis factor, and transforming growth factor were also significantly altered by PD123319. These data suggest that AT2 blockade is involved in significant changes in myocardial extracellular matrix components translating into decreases in tissue mass in the salt-sensitive ANP−/− animal.

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