scholarly journals Transcriptomic and proteomic analysis of putative digestive proteases in the salivary gland and gut of Empoasca (Matsumurasca) onukii Matsuda

2020 ◽  
Author(s):  
Ensi Shao ◽  
Yujuan Song ◽  
Yaomin Wang ◽  
Yichen Liao ◽  
Yufei Luo ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Enzymatic Activity ◽  
Serine Proteases ◽  
Cathepsin L ◽  
Transcript Abundance ◽  
Nilaparvata Lugens ◽  
Integrated Analysis ◽  
Abundance Pattern ◽  
Digestive Proteases ◽  
Tea Plants

Abstract Background: Infestation by tea green leafhoppers, Empoasca (Matsumurasca) onukii , could cause a series of biochemical changes in tea leaves. As a typical cell-rupture feeder, E. onukii secretes proteases while probing with its stylet into the tender shoots of tea plants ( Camellia sinensis ). This study identified and analyzed proteases specifically expressed in the salivary gland (SG) and gut of E. onukii through enzymatic activity assays, complemented with an integrated analysis of transcriptome and proteome data.Results: In total, 129 contigs representing seven types of putative proteases were identified. Transcript abundance of digestive proteases and enzymatic activity assays showed that cathepsin B-like protease, cathepsin L-like protease, and serine proteases (trypsin- and chymotrypsin-like protease) were highly abundant in the gut while moderately abundant in the SG. The abundance pattern of digestive proteases in the SG and gut of E. onukii differed from that of other hemipterans including Nilaparvata lugens , Laodelphax striatellus , Acyrthosiphum pisum , Halyomorpha halys and Nephotettix cincticeps . Phylogenetic analysis showed that aminopeptidase N-like proteins and serine proteases abundant in the SG or gut of hemipterans were distributed to two distinct clusters.Conclusions: Altogether, this study provide insightful information on the digestive system of E. onukii and observed different patterns of proteases abundant in the SG and gut of E. onukii , in comparison with other five hemipteran species. These results will be beneficial in understanding the interaction between tea plants and E. onukii .

scholarly journals Transcriptomic and proteomic analysis of putative digestive proteases in the salivary gland and gut of Empoasca(Matsumurasca) onukii Matsuda

2020 ◽  
Author(s):  
Ensi Shao ◽  
Yujuan Song ◽  
Yaomin Wang ◽  
Yichen Liao ◽  
Yufei Luo ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Enzymatic Activity ◽  
Serine Proteases ◽  
Cathepsin L ◽  
Transcript Abundance ◽  
Nilaparvata Lugens ◽  
Integrated Analysis ◽  
Abundance Pattern ◽  
Digestive Proteases ◽  
Tea Plants

Abstract Background: Infestation by tea green leafhoppers, Empoasca (Matsumurasca) onukii, could cause a series of biochemical changes in tea leaves. As a typical cell-rupture feeder, E. onukii secretes proteases while probing with its stylet into the tender shoots of tea plants (Camellia sinensis). This study identified and analyzed proteases specifically expressed in the salivary gland (SG) and gut of E. onukii through enzymatic activity assays, complemented with an integrated analysis of transcriptome and proteome data.Results: In total, 129 contigs representing seven types of putative proteases were identified. Transcript abundance of digestive proteases and enzymatic activity assays showed that cathepsin B, cathepsin L, and serine proteases (trypsin and chymotrypsin) were highly abundant in the gut while moderately abundant in the SG. The abundance pattern of digestive proteases in the SG and gut of E. onukii differed from that of other hemipterans including Nilaparvata lugens, Laodelphax striatellus, Nephotettix cincticeps and Acyrthosiphum pisum. Phylogenetic analysis showed that aminopeptidase N-like proteins and serine proteases abundant in the SG or gut of hemipterans were distributed to two distinct clusters. Conclusions: Altogether, this study provide insightful information on the digestive system of E. onukii and observed different patterns of proteases abundant in the SG and gut of E. onukii, in comparison with other five hemipteran species. These results will be beneficial in understanding the interaction between tea plants and E. onukii.

scholarly journals Transcriptomic and proteomic analysis of putative digestive proteases in the salivary gland and gut of Empoasca (Matsumurasca) onukii Matsuda

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Ensi Shao ◽  
Yujuan Song ◽  
Yaomin Wang ◽  
Yichen Liao ◽  
Yufei Luo ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Enzymatic Activity ◽  
Serine Proteases ◽  
Cathepsin L ◽  
Transcript Abundance ◽  
Nilaparvata Lugens ◽  
Integrated Analysis ◽  
Abundance Pattern ◽  
Digestive Proteases ◽  
Tea Plants

Abstract Background Infestation by tea green leafhoppers (Empoasca (Matsumurasca) onukii) can cause a series of biochemical changes in tea leaves. As a typical cell-rupture feeder, E. onukii secretes proteases while using its stylet to probe the tender shoots of tea plants (Camellia sinensis). This study identified and analyzed proteases expressed specifically in the salivary gland (SG) and gut of E. onukii through enzymatic activity assays complemented with an integrated analysis of transcriptomic and proteomic data. Results In total, 129 contigs representing seven types of putative proteases were identified. Transcript abundance of digestive proteases and enzymatic activity assays showed that cathepsin B-like protease, cathepsin L-like protease, and serine proteases (trypsin- and chymotrypsin-like protease) were highly abundant in the gut but moderately abundant in the SG. The abundance pattern of digestive proteases in the SG and gut of E. onukii differed from that of other hemipterans, including Nilaparvata lugens, Laodelphax striatellus, Acyrthosiphum pisum, Halyomorpha halys and Nephotettix cincticeps. Phylogenetic analysis showed that aminopeptidase N-like proteins and serine proteases abundant in the SG or gut of hemipterans formed two distinct clusters. Conclusions Altogether, this study provides insightful information on the digestive system of E. onukii. Compared to five other hemipteran species, we observed different patterns of proteases abundant in the SG and gut of E. onukii. These results will be beneficial in understanding the interaction between tea plants and E. onukii.

scholarly journals Molecular and Biochemical Analysis of Periplastidial Starch Metabolism in the Cryptophyte Guillardia theta

2006 ◽  
Vol 5 (6) ◽  
pp. 964-971 ◽  
Author(s):  
Ilka Haferkamp ◽  
Philippe Deschamps ◽  
Michelle Ast ◽  
Wolfgang Jeblick ◽  
Uwe Maier ◽  
...  
Keyword(s):  
Continuous Light ◽  
Subcellular Location ◽  
Transcript Abundance ◽  
Transport Processes ◽  
Phosphate Transporter ◽  
Light Phase ◽  
Abundance Pattern ◽  
Total Enzyme Activity ◽  
Triose Phosphate ◽  
Guillardia Theta

ABSTRACT Starch in synchronously grown Guillardia theta cells accumulates throughout the light phase, followed by a linear degradation during the night. In contrast to the case for other unicellular algae such as Chlamydomonas reinhardtii, no starch turnover occurred in this organism under continuous light. The gene encoding granule-bound starch synthase (GBSS1), the enzyme responsible for amylose synthesis, displays a diurnal expression cycle. The pattern consisted of a maximal transcript abundance around the middle of the light phase and a very low level during the night. This diurnal regulation of GBSS1 transcript abundance was demonstrated to be independent of the circadian clock but tightly light regulated. A similar yet opposite type of regulation pattern was found for two α-amylase isoforms and for one of the two plastidic triose phosphate transporter genes investigated. In these cases, however, the transcript abundance peaked in the night phase. The second plastidic triose phosphate transporter gene had the GBSS1 mRNA abundance pattern. Quantification of the GBSS1 activity revealed that not only gene expression but also total enzyme activity exhibited a maximum in the middle of the light phase. To gain a first insight into the transport processes involved in starch biosynthesis in cryptophytes, we demonstrated the presence of both plastidic triose phosphate transporter and plastidic ATP/ADP transporter activities in proteoliposomes harboring either total membranes or plastid envelope membranes from G. theta. These molecular and biochemical data are discussed with respect to the environmental conditions experienced by G. theta and with respect to the unique subcellular location of starch in cryptophytes.

scholarly journals Putative Serine Protease Effectors of Clavibacter michiganensis Induce a Hypersensitive Response in the Apoplast of Nicotiana Species

2015 ◽  
Vol 28 (11) ◽  
pp. 1216-1226 ◽  
Author(s):  
You Lu ◽  
Noriyuki Hatsugai ◽  
Fumiaki Katagiri ◽  
Carol A. Ishimaru ◽  
Jane Glazebrook
Keyword(s):  
Nicotiana Tabacum ◽  
Enzymatic Activity ◽  
Hypersensitive Response ◽  
Serine Proteases ◽  
Host Plants ◽  
Ectopic Expression ◽  
Related Protein ◽  
Clavibacter Michiganensis ◽  
Serine Residue ◽  
Nicotiana Species

Clavibacter michiganensis subspp. michiganensis and sepedonicus cause diseases on solanaceous crops. The genomes of both subspecies encode members of the pat-1 family of putative serine proteases known to function in virulence on host plants and induction of hypersensitive responses (HR) on nonhosts. One gene of this family in C. michiganensis subsp. sepedonicus, chp-7, is required for triggering HR in Nicotiana tabacum. Here, further investigation revealed that mutation of the putative catalytic serine residue at position 232 to threonine abolished the HR induction activity of Chp-7, suggesting that enzymatic activity is required. Purified Chp-7 triggered an HR in N. tabacum leaves in the absence of the pathogen, indicating Chp-7 itself is the HR elicitor from C. michiganensis subsp. sepedonicus. Ectopic expression of chp-7 constructs in N. tabacum leaves revealed that Chp-7 targeted to the apoplast triggered an HR while cytoplasmic Chp-7 did not, indicating that Chp-7 induces the HR in the apoplast of N. tabacum leaves. Chp-7 also induced HR in N. sylvestris, a progenitor of N. tabacum, but not in other Nicotiana species tested. ChpG, a related protein from C. michiganensis subsp. michiganensis, also triggered HR in N. tabacum and N. sylvestris. Unlike Chp-7, ChpG triggered HR in N. clevelandii and N. glutinosa.

scholarly journals Identification and analysis of miRNAs in IR56 rice in response to BPH infestations of different virulence levels

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Satyabrata Nanda ◽  
San-Yue Yuan ◽  
Feng-Xia Lai ◽  
Wei-Xia Wang ◽  
Qiang Fu ◽  
...  
Keyword(s):  
Regulatory Network ◽  
Molecular Mechanisms ◽  
Brown Planthopper ◽  
Enrichment Analysis ◽  
Transcript Abundance ◽  
Defense Responses ◽  
Nilaparvata Lugens ◽  
Mirna Sequence ◽  
Rna Profiling ◽  
Underlying Mechanisms

Abstract Rice production and sustainability are challenged by its most dreadful pest, the brown planthopper (Nilaparvata lugens Stål, BPH). Therefore, the studies on rice-BPH interactions and their underlying mechanisms are of high interest. The rice ontogenetic defense, such as the role of microRNAs (miRNAs) has mostly been investigated against the pathogens, with only a few reports existing against the insect infestations. Thus, revealing the involvement of rice miRNAs in response to BPH infestations will be beneficial in understanding these complex interactions. In this study, the small RNA profiling of the IR56 rice in response to separate BPH infestations of varied virulence levels identified the BPH-responsive miRNAs and revealed the differential transcript abundance of several miRNAs during a compatible and incompatible rice-BPH interaction. The miRNA sequence analysis identified 218 known and 28 novel miRNAs distributed in 54 miRNA families. Additionally, 138 and 140 numbers of differentially expressed (DE) miRNAs were identified during the compatible and incompatible interaction, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed the target gene candidates of DE miRNAs (including osa-miR2871a-3p, osa-miR172a, osa-miR166a-5p, osa-miR2120, and osa-miR1859) that might be involved in the IR56 rice defense responses against BPH infestation. Conversely, osa-miR530-5p, osa-miR812s, osa-miR2118g, osa-miR156l-5p, osa-miR435 and two of the novel miRNAs, including novel_16 and novel_52 might negatively modulate the IR56 rice defense. The expressional validation of the selected miRNAs and their targets further supported the IR56 rice defense regulatory network. Based on our results, we have proposed a conceptual model depicting the miRNA defense regulatory network in the IR56 rice against BPH infestation. The findings from the study add further insights into the molecular mechanisms of rice-BPH interactions and will be helpful for the future researches.

The serpin proteinase inhibitor 8: An endogenous furin inhibitor released from human platelets

2006 ◽  
Vol 95 (02) ◽  
pp. 243-252 ◽  
Author(s):  
Julie Leblond ◽  
Marie-Hélène Laprise ◽  
Simon Gaudreau ◽  
Francine Grondin ◽  
Walter Kisiel ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Proteinase Inhibitor ◽  
Serine Proteases ◽  
Antithrombin Iii ◽  
Direct Interaction ◽  
Human Platelets ◽  
Stable Complex ◽  
Functional Responses ◽  
Pai 1 ◽  
Platelet Functions

SummaryThe cornerstone of hemostasis is the ability of the organism to limit the enzymatic processes involved, thereby avoiding thrombosis. For this, anticoagulant systems in place involve serpins, such as PAI-1 and antithrombin III, which bind to their targeted serine proteases and limit their period of activity. We have previously identified the serine protease furin as a platelet-derived enzyme with an intrinsic role in platelet functions. We now report that furin enzymatic activity decreased rapidly following platelet activation, corresponding with the increase in formation of a high 180 M r SDS-stable complex composed of furin and the PI8 serpin. PI8 is shown to be a platelet-derived constituent, synthesized by megakaryocytes and stored in platelets prior to its release. Immunoprecipitation and purification of the PI8-furin complex confirmed their direct interaction and indicates that one of the roles of PI8 is to inhibit furin enzymatic activity. Furthermore, our findings demonstrate the inhibitory capacity of exogenous PI8 in platelet aggregation assays. The finding that PI8 is released by platelets and controls functional responses suggests a role for this serpin in platelet-regulated pathophysiological responses.

scholarly journals Integrated analysis and transcript abundance modelling of H3K4me3 and H3K27me3 in developing secondary xylem

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Steven G. Hussey ◽  
Mattheus T. Loots ◽  
Karen van der Merwe ◽  
Eshchar Mizrachi ◽  
Alexander A. Myburg
Keyword(s):  
Secondary Xylem ◽  
Transcript Abundance ◽  
Integrated Analysis

Procongopain from Trypanosoma congolense Is Processed at Basic pH: An Unusual Feature among Cathepsin L-Like Cysteine Proteases

2003 ◽  
Vol 384 (6) ◽  
pp. 921-927 ◽  
Author(s):  
C. Serveau ◽  
A. Boulangé ◽  
F. Lecaille ◽  
F. Gauthier ◽  
E. Authié ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Dextran Sulfate ◽  
Cathepsin L ◽  
Active Form ◽  
Fold Increase ◽  
Cysteine Proteases ◽  
Trypanosoma Congolense ◽  
Unusual Feature ◽  
One Step ◽  
Host Parasite

Abstract Congopain, the major cysteine protease from Trypanosoma congolense, is synthesized as an inactive zymogen, and further converted into its active form after removal of the proregion, most probably via an autocatalytic mechanism. Processing of recombinant procongopain occurs via an apparent one-step or a multistep mechanism depending on the ionic strength. The auto-activation is pH-dependent, with an optimum at pH 4.0, and no activation observed at pH 6.0. After addition of dextran sulfate (10 ug/ml), an approx. 20-fold increase of processing (expressed as enzymatic activity) is observed. Furthermore, in the presence of dextran sulfate, procongopain can be processed at pH 8.0, an unusual feature among papainlike enzymes. Detection of procongopain and trypanosomal enzymatic activity in the plasma of T. congolenseinfected cattle, together with the capacity of procongopain to be activated at weakly basic pH, suggest that procongopain may be extracellularly processed in the presence of blood vessel glycosaminoglycans, supporting the hypothesis that congopain acts as a pathogenic factor in host-parasite relationships.

scholarly journals Effects of Local and Systemic Immune Challenges on the Expression of Selected Salivary Genes in the Malaria Mosquito Anopheles coluzzii

Pathogens ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1300
Author(s):  
Giulia Bevivino ◽  
Bruno Arcà ◽  
Fabrizio Lombardo
Keyword(s):  
Salivary Gland ◽  
Salivary Glands ◽  
Antimicrobial Properties ◽  
Transcript Abundance ◽  
Structural Features ◽  
Anopheles Coluzzii ◽  
Transcriptional Modulation ◽  
Immune Challenges

Salivary glands play a crucial tripartite role in mosquito physiology. First, they secrete factors that greatly facilitate both sugar and blood meal acquisition. Second, the transmission of pathogens (parasites, bacteria and viruses) to the vertebrate host requires both the recognition and invasion of the salivary glands. Third, they produce immune factors that both protect the organ from invading pathogens and are also able to exert their activity in the crop and the midgut when saliva is re-ingested during feeding. Studies on mosquito sialomes have revealed the presence of several female and/or male salivary gland-specific or enriched genes whose function is completely unknown so far. We focused our attention on these orphan genes, and we selected, according to sequence and structural features, a shortlist of 11 candidates with potential antimicrobial properties. Afterwards, using qPCR, we investigated their expression profile at 5 and 24 h after an infectious sugar meal (local challenge) or thoracic microinjection (systemic challenge) of Gram-negative (Escherichia coli, EC) or Gram-positive (Staphylococcus aureus, SA) bacteria. We observed a general increase in the transcript abundance of our salivary candidates between 5 and 24 h after local challenge. Moreover, transcriptional modulation was determined by the nature of the stimulus, with salivary gland-enriched genes (especially hyp15 upon SA stimulus) upregulated shortly after the local challenge and later after the systemic challenge. Overall, this work provides one of the first contributions to the understanding of the immune role of mosquito salivary glands. Further characterization of salivary candidates whose expression is modulated by immune challenge may help in the identification of possible novel antimicrobial peptides.

scholarly journals Partial characterization of digestive proteases in Pacific red snapper Lutjanus peru Nichols & Murphy, 1922 (Perciformes: Lutjanidae)

2021 ◽  
Vol 49 (3) ◽  
pp. 442-450
Author(s):  
Emyr S. Peña-Marín ◽  
Leonardo Ibarra-Castro ◽  
Juan M. Martínez-Brown ◽  
Iris A. Hernández-López ◽  
Dariel Tovar-Ramírez ◽  
...  
Keyword(s):  
Serine Proteases ◽  
Acid Protease ◽  
Red Snapper ◽  
Alkaline Proteases ◽  
Digestive Proteases ◽  
Optimum Ph ◽  
Commercial Species ◽  
Electrophoretic Techniques ◽  
Pepstatin A

Pacific red snapper (Lutjanus peru) is an important commercial species in Mexico with great aquaculture potential; however, digestive physiology is still unknown. Therefore, the objective of the present work was to characterize the digestive proteases of L. peru juvenile using biochemical and electrophoretic techniques. Results showed a higher acid protease activity than the alkaline proteases, trypsin, chymotrypsin, and leucine aminopeptidase (LAP). The optimum temperature for acid proteases was between 30 to 40°C. Trypsin activity showed two maximum peaks of temperature (30 and 50°C), while alkaline proteases, chymotrypsin, and LAP had optimum temperatures of 50, 50 to 60, and 40°C, respectively. Moreover, the optimum pH of acid proteases was between 2 and 3. Also, alkaline proteases, trypsin, chymotrypsin showed pH optimums at pH 6, 9, and 5, respectively, although LAP showed two optimum pH values at 6 and 9. Acid protease zymogram showed three isoforms, totally inhibited by pepstatin A. Alkaline protease zymogram revealed six bands (125.4, 67.2, 57.9, 48.6, 29.8, and 26.9 kDa), which were inhibited by specific serine-proteases and metalloproteases inhibitors. In conclusion, the main digestion in L. peru depends on stomach proteases, which are characteristic of carnivorous fish, followed by intestinal digestion supported mainly by chymotrypsin.

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