Expression Pattern Of Proteases Research Articles

Expression of macromolecular organic nitrogen degrading enzymes identifies potential mediators of soil organic N availability to an annual grass

Nitrogen (N) is frequently limiting to plant growth, in part because most soil N is present as polymeric organic compounds that are not readily taken up by plants. Microbial depolymerization of these large macromolecular N-substrates gradually releases available inorganic N. While many studies have researched and modeled controls on soil organic matter formation and bulk N mineralization, the ecological—spatial, temporal and phylogenetic—patterns underlying organic N degradation remain unclear. We analyzed 48 time-resolved metatranscriptomes and quantified N-depolymerization gene expression to resolve differential expression by soil habitat and time in specific taxonomic groups and gene-based guilds. We observed much higher expression of extracellular serine-type proteases than other extracellular N-degrading enzymes, with protease expression of predatory bacteria declining with time and other taxonomic patterns driven by the presence (Gammaproteobacteria) or absence (Thermoproteota) of live roots and root detritus (Deltaproteobacteria and Fungi). The primary chitinase chit1 gene was more highly expressed by eukaryotes near root detritus, suggesting predation of fungi. In some lineages, increased gene expression over time suggests increased competitiveness with rhizosphere age (Chloroflexi). Phylotypes from some genera had protease expression patterns that could benefit plant N nutrition, for example, we identified a Janthinobacterium phylotype and two Burkholderiales that depolymerize organic N near young roots and a Rhizobacter with elevated protease levels near mature roots. These taxon-resolved gene expression results provide an ecological read-out of microbial interactions and controls on N dynamics in specific soil microhabitats and could be used to target potential plant N bioaugmentation strategies.

Human Melanoma-Associated Mast Cells Display a Distinct Transcriptional Signature Characterized by an Upregulation of the Complement Component 3 That Correlates With Poor Prognosis.

Cutaneous melanoma is one of the most aggressive human malignancies and shows increasing incidence. Mast cells (MCs), long-lived tissue-resident cells that are particularly abundant in human skin where they regulate both innate and adaptive immunity, are associated with melanoma stroma (MAMCs). Thus, MAMCs could impact melanoma development, progression, and metastasis by secreting proteases, pro-angiogenic factors, and both pro-inflammatory and immuno-inhibitory mediators. To interrogate the as-yet poorly characterized role of human MAMCs, we have purified MCs from melanoma skin biopsies and performed RNA-seq analysis. Here, we demonstrate that MAMCs display a unique transcriptome signature defined by the downregulation of the FcεRI signaling pathway, a distinct expression pattern of proteases and pro-angiogenic factors, and a profound upregulation of complement component C3. Furthermore, in melanoma tissue, we observe a significantly increased number of C3+ MCs in stage IV melanoma. Moreover, in patients, C3 expression significantly correlates with the MC-specific marker TPSAB1, and the high expression of both markers is linked with poorer melanoma survival. In vitro, we show that melanoma cell supernatants and tumor microenvironment (TME) mediators such as TGF-β, IL-33, and IL-1β induce some of the changes found in MAMCs and significantly modulate C3 expression and activity in MCs. Taken together, these data suggest that melanoma-secreted cytokines such as TGF-β and IL-1β contribute to the melanoma microenvironment by upregulating C3 expression in MAMCs, thus inducing an MC phenotype switch that negatively impacts melanoma prognosis.

Heat-Stable Antifungal Factor and Expression Patterns of Protease by Chitinase-Producing Bacterium, Pedobacter sp. PR-M6

This study was conducted to investigate the protease expression patterns of the novel chitinase-producing bacterium Pedobacter sp. PR-M6. Protease were detected on SDS-PAGE gel as condition of 4℃, 25℃ and 37℃ to activate of enzymes obtained from incubation of LB, NB and 1% skim milk medium at 4℃, 20℃ and 30℃ after 21 days of incubation, respectively. Pedobacter sp. PR-M6 displayed proteolytic activity on SDS-PAGE gels containing 0.1% (w/v) gelatin, and protease isozymes of PR-M6 were expressed as two major bands (P7 and P8) and seven minor bands (P1, P2, P3, P4, P5, P6, and P9). Protease isozymes of PR-M6 were inhibited at 4℃ and 25℃. The two protease isozymes P5 and P6 were newly expressed in the LB culture medium at 20℃. To investigate the properties of a heat-stable antifungal factor (HSAF), PR-M6 stain was cultured to obtain the products of biosynthesis for incubation in LB medium. HSAF exhibited active biosynthesis at 20℃ for incubation in LB medium. Crude HASF obtained from LB20 medium showed hyphal growth inhibition of Rhizoctonia solani. Inhibition rate of R. solani growth was as high as 67% at 24 h and 47.8% at 48 h after incubation with 1,000 ppm of crude HSAF. Trypan blue stained the inside of the edge for destructed R. solani cell walls after crude HSAF treatment. These results indicated that HSAF and protease from Pedobacter sp. PR-M6 could be used as biocontrol agents against phytopathogens in agriculture.

The Effect of Salinity on Growth, Antagonistic Potential, Protease Activity, and Proline Content of Trichoderma harzianum

Salinity is one of the major limiting factors for sustainable crop production and the growth of beneficial microorganisms. Screening of biocontrol agent Trichoderma harzianum isolates under saline conditions could be necessary to select more efficient strains to alleviate salt stress through biopriming in plants. The present study aimed to investigate the antagonistic capacity of 14 T. harzianum isolates against Macrophomina phaseolina as well as the growth, protease activity, and proline content under different concentrations of salt. All isolates except Th15 and Th18 exhibited an over 85% inhibition against M. phaseolina at 0 mM NaCl in c and showed a noticeable decline in their antagonistic capacity as salt concentration increased. Colony growth decreased with increasing salt concentrations in all isolates tested. The growth of all isolates at 0 mM was significantly higher than the other NaCl treatments except at 70 mM NaCl. Protease activity also declined with the increased level of salt. Isolates displayed a wide range of protease expression patterns even at the same salinity as in the case of proline content. Unlike protease enzyme, proline content significantly increased at 240 mM NaCl. Salinity played a significant role in T. harzianum isolates with regards to the growth, antagonistic capacity, protease activity, and proline content.

Expression profile of the Schistosoma japonicum degradome reveals differential protease expression patterns and potential anti-schistosomal intervention targets.

Blood fluke proteases play pivotal roles in the processes of invasion, nutrition acquisition, immune evasion, and other host-parasite interactions. Hundreds of genes encoding putative proteases have been identified in the recently published schistosome genomes. However, the expression profiles of these proteases in Schistosoma species have not yet been systematically analyzed. We retrieved and culled the redundant protease sequences of Schistosoma japonicum, Schistosoma mansoni, Echinococcus multilocularis, and Clonorchis sinensis from public databases utilizing bioinformatic approaches. The degradomes of the four parasitic organisms and Homo sapiens were then comparatively analyzed. A total of 262 S. japonicum protease sequences were obtained and the expression profiles generated using whole-genome microarray. Four main clusters of protease genes with different expression patterns were identified: proteases up-regulated in hepatic schistosomula and adult worms, egg-specific or predominantly expressed proteases, cercaria-specific or predominantly expressed proteases, and constantly expressed proteases. A subset of protease genes with different expression patterns were further validated using real-time quantitative PCR. The present study represents the most comprehensive analysis of a degradome in Schistosoma species to date. These results provide a firm foundation for future research on the specific function(s) of individual proteases and may help to refine anti-proteolytic strategies in blood flukes.

Differential protease activity augments polyphagy in Helicoverpa armigera

Helicoverpa armigera (Lepidoptera: Noctuidae) and other polyphagous agricultural pests are extending their plant host range and emerging as serious agents in restraining crop productivity. Dynamic regulation, coupled with a diversity of digestive and detoxifying enzymes, play a crucial role in the adaptation of polyphagous insects. To investigate the functional intricacy of serine proteases in the development and polyphagy of H. armigera, we profiled the expression of eight trypsin-like and four chymotrypsin-like phylogenetically diverse mRNAs from different life stages of H. armigera reared on nutritionally distinct host plants. These analyses revealed diet- and stage-specific protease expression patterns. The trypsins expressed showed structural variations, which might result in differential substrate specificity and interaction with inhibitors. Protease profiles in the presence of inhibitors and their mass spectrometric analyses revealed insight into their differential activity. These findings emphasize the differential expression of serine proteases and their consequences for digestive physiology in promoting polyphagy in H. armigera.

Disease-specific expression patterns of proteases in synovial tissues

To assess whether protease expression patterns can be discriminated according to matrix degradation mechanisms in aseptic prosthesis loosening (APL), rheumatoid arthritis (RA), and osteoarthritis (OA), we immunohistochemically examined the expressions of matrix metalloproteinase-1 and cathepsins B, D, and L in periprosthetic synovial-like interface tissues from 32 patients with failed prosthetic hips, from 29 RA-patients with hip synovial membranes, and from 35 patients with primary OA. Numerical values, calculated for the positivity of each protease, were used to rank the staining patterns, and a multivariate analysis was carried out to examine the discriminant probabilities. As a result of stepwise linear discriminant analyses, the three groups were successfully discriminated with probabilities of 100%, 62.1%, and 77.1%, respectively. Cathepsin L was significantly related to the discrimination of APL from RA and primary OA. Disease-specific protease activation pathways might exist, and cathepsin L can be a key enzyme for APL pathogenesis. Level of evidence: Prognostic study, level III (retrospective study).

Mast cell numbers and protease expression patterns in biopsy specimens following renal transplantation from living‐related donors predict long‐term graft function

In human kidney transplantation the main cause of declining long-term graft function is chronic allograft nephropathy (CAN). Recent studies have implicated human mast cells (MC) in chronic inflammation and fibrosis, MC can be subtyped according to protease content: MC(T) containing tryptase only and MC(TC) containing both tryptase and chymase. We investigated immunohistochemically whether numbers and subtypes of MC in biopsy specimens 100 d after transplantation could predict subsequent fibrosis and graft dysfunction. The total number of MC/high-power field at 100 d after transplantation correlated significantly with change in creatinine clearance (DeltaCcr), defined as (Ccr at 100 d) - (Ccr at 3 yr) (R = 0.597, p = 0.0021); fibrosis index (FI) at 100 d (R = 0.583, p = 0.0066); and DeltaFI, defined as (FI at 3 yr) - (FI at 100 d) (R = 0.406, p < 0.05). The ratio of MC(TC) to total MC at 100 d also correlated with DeltaCcr (R = 0.491, p = 0.0148), FI at 100 d (R = 0.527, p = 0.0081), and DeltaFI (R = 0.417, p < 0.05). Thus, increases in number of total MC and the ratio of MC(TC) to total MC in early biopsy specimens were related to decline of long-term graft function and fibrosis.

Leukemia inhibitory factor triggers activation of signal transducer and activator of transcription 3, proliferation, invasiveness, and altered protease expression in choriocarcinoma cells

Extravillous trophoblast cells resemble cancer cells with regard to their intrinsic invasiveness. They invade decidual tissue, but, unlike tumor cells, shut down their invasive properties, when they become inappropriate. Stimuli involved in the modulation of invasion, as well as their underlying signaling mechanisms require further clarification. We were especially interested in discovering signals capable of stimulating invasion in otherwise low-invasive cells involved in reproduction. Using the choriocarcinoma cell line Jeg-3 as a model, we have addressed the potential role of cytokine/growth factor-driven activation of signal transducer and activator of transcription 3 (STAT3) in this process. Jeg-3 cells were treated with various factors known to induce trophoblast proliferation, differentiation, migration, or invasiveness (insulin-like-growth-factor-II (IGF-II), hepatocyte growth factor (HGF), interleukin-6 (IL-6), and leukemia inhibitory factor (LIF)). Only LIF elicited strong tyrosine phosphorylation and specific DNA-binding activity of STAT3. It induced a significant acceleration of cell proliferation and promoted the capability of Jeg-3 cells to invade into an artificial extracellular matrix. Moreover, LIF influenced the expression pattern of proteases and protease inhibitors with potential relevance for invasiveness (downregulation of mRNA for tissue inhibitor of metalloproteinase 1 (TIMP-1) and upregulation of mRNA for caspase-4). In conjunction with earlier work, in which we found that STAT3 DNA-binding activity was increased in invasive cells (choriocarcinoma, first trimester trophoblasts) and absent in non-invasive cells (term trophoblasts), these findings suggest a connection between LIF-driven STAT3 activity and invasiveness of choriocarcinoma and trophoblast cells.

1870: Mast Cell Numbers and Protease Expression Patterns in Biopsy Specimens Following Renal Transplantation From Living Related Donors Predict Long-Term Graft Function

1870: Mast Cell Numbers and Protease Expression Patterns in Biopsy Specimens Following Renal Transplantation From Living Related Donors Predict Long-Term Graft Function

Expression pattern and functional studies of matrix degrading proteases and their inhibitors in the mouse corpus luteum

The formation of the corpus luteum (CL) is accompanied with angiogenesis and tissue remodeling and its regression involves tissue degradation. Matrix degrading proteases such as plasminogen activators (PAs) and matrix metalloproteinases (MMPs) are thought to play important roles in such controlled proteolytic processes. In this study, in situ hybridization has been used to examine the regulation and expression pattern of mRNAs coding for proteases and protease inhibitors belonging to the PA- and MMP-systems during the life cycle of the CL in an adult pseudopregnant mouse model. Of the nine proteases and five protease inhibitors that were studied, the majority were found to be temporally expressed during the formation and/or the regression of the CL. However, the mRNAs coding for urokinase type PA (uPA), membrane-type 1 MMP (MT1-MMP), and tissue inhibitor of metalloproteinases type-3 (TIMP-3) were constantly expressed in the mouse CL throughout its whole life span. To study the functional role of uPA in the CL, we analyzed luteal formation and function in uPA deficient mice. Our results revealed no significant difference in ovarian weight, serum progesterone levels, and blood vessel density in the functional CL between uPA deficient and wild type control mice. The temporal and spatial expression pattern of proteases and protease inhibitors during the CL life span suggests that members of the PA- and MMP-systems may play important roles in the angiogenesis and tissue remodeling processes during CL formation, as well as in the tissue degradation during luteal regression. However, the absence of reproductive phenotypes in mice lacking uPA and several other matrix degrading proteases indicates that there are redundancies among different matrix degrading proteases or that tissue remodeling in the ovary may involve other additional unique elements.

Studies on phenotypic expression patterns of proteases and protease inhibitors in benign compared to malignant prostate tissues.

Studies on phenotypic expression patterns of proteases and protease inhibitors in benign compared to malignant prostate tissues

Overexpression of cathepsin B and urokinase plasminogen activator is associated with increased risk of recurrence and metastasis in patients with chondrosarcoma

Deregulation of the cellular protease network has been shown to be responsible for aggressive clinical behavior in several common human malignancies. In the current study, the authors evaluated the expression patterns of proteases in patients with chondrosarcoma of bone and correlated these patterns with clinical outcome. The expression levels of urokinase plasminogen activator; matrix metalloproteinase types-1, -2, and -9; and cathepsins B and L were determined immunohistochemically in 114 cases of chondrosarcomas of bone and were correlated with their clinicopathologic parameters as well as with long term follow-up data. Overexpression of cathepsin B was associated with a high rate of local recurrence (P = 0.006) and a decreased recurrence free survival (P = 0.005). Overexpression of urokinase plasminogen activator was associated with an increased rate of metastasis (P = 0. 013), a decreased metastasis free survival (P = 0.016), and a decreased 5-year overall survival rate (P = 0.048). The univariate Cox model showed that tumor extension into soft tissue, high histologic grade, and overexpression of cathepsin B were predictors of adverse outcome. Multivariate analysis showed only overexpression of cathepsin B and tumor extension into soft tissue to be independent predictors of local recurrence. Overexpression of cathepsin B and urokinase plasminogen activator can be used to identify those patients with chondrosarcoma of bone who have an increased risk of local recurrence and distant metastases.