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Therapeutic Methods and Therapies TCIM
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2.
BMC Genomics ; 21(1): 331, 2020 Apr 29.
Article in English | MEDLINE | ID: mdl-32349672

ABSTRACT

BACKGROUND: Salivary cell secretion (SCS) plays a critical role in blood feeding by medicinal leeches, making them of use for certain medical purposes even today. RESULTS: We annotated the Hirudo medicinalis genome and performed RNA-seq on salivary cells isolated from three closely related leech species, H. medicinalis, Hirudo orientalis, and Hirudo verbana. Differential expression analysis verified by proteomics identified salivary cell-specific gene expression, many of which encode previously unknown salivary components. However, the genes encoding known anticoagulants have been found to be expressed not only in salivary cells. The function-related analysis of the unique salivary cell genes enabled an update of the concept of interactions between salivary proteins and components of haemostasis. CONCLUSIONS: Here we report a genome draft of Hirudo medicinalis and describe identification of novel salivary proteins and new homologs of genes encoding known anticoagulants in transcriptomes of three medicinal leech species. Our data provide new insights in genetics of blood-feeding lifestyle in leeches.


Subject(s)
Genome , Hirudo medicinalis/genetics , Salivary Proteins and Peptides/genetics , Animals , Anticoagulants/metabolism , Gene Expression Profiling , Gene Expression Regulation , Hirudo medicinalis/metabolism , Leeches/classification , Leeches/genetics , Leeches/metabolism , Proteomics , Saliva/metabolism , Salivary Proteins and Peptides/metabolism
3.
BMC Plant Biol ; 19(1): 9, 2019 Jan 07.
Article in English | MEDLINE | ID: mdl-30616513

ABSTRACT

BACKGROUND: Cryptic peptides (cryptides) are small bioactive molecules generated via degradation of functionally active proteins. Only a few examples of plant cryptides playing an important role in plant defense have been reported to date, hence our knowledge about cryptic signals hidden in protein structure remains very limited. Moreover, little is known about how stress conditions influence the size of endogenous peptide pools, and which of these peptides themselves have biological functions is currently unclear. RESULTS: Here, we used mass spectrometry to comprehensively analyze the endogenous peptide pools generated from functionally active proteins inside the cell and in the secretome from the model plant Physcomitrella patens. Overall, we identified approximately 4,000 intracellular and approximately 500 secreted peptides. We found that the secretome and cellular peptidomes did not show significant overlap and that respective protein precursors have very different protein degradation patterns. We showed that treatment with the plant stress hormone methyl jasmonate induced specific proteolysis of new functional proteins and the release of bioactive peptides having an antimicrobial activity and capable to elicit the expression of plant defense genes. Finally, we showed that the inhibition of protease activity during methyl jasmonate treatment decreased the secretome antimicrobial potential, suggesting an important role of peptides released from proteins in immune response. CONCLUSIONS: Using mass-spectrometry, in vitro experiments and bioinformatics analysis, we found that methyl jasmonate acid induces significant changes in the peptide pools and that some of the resulting peptides possess antimicrobial and regulatory activities. Moreover, our study provides a list of peptides for further study of potential plant cryptides.


Subject(s)
Acetates/pharmacology , Anti-Infective Agents/metabolism , Bryopsida/metabolism , Cyclopentanes/pharmacology , Oxylipins/pharmacology , Peptides/metabolism , Plant Growth Regulators/pharmacology , Anti-Infective Agents/isolation & purification , Bacillus subtilis/drug effects , Bryopsida/drug effects , Escherichia coli/drug effects , Mass Spectrometry , Microbial Sensitivity Tests , Peptides/isolation & purification
4.
Sci Rep ; 7(1): 11330, 2017 09 12.
Article in English | MEDLINE | ID: mdl-28900116

ABSTRACT

Despite the fact the term "proteome" was proposed to characterize a set of proteins in one of mycoplasma species, proteome response to various exposures in this bacteria are still obscure. Commonly, authors studying proteomic response on perturbation models in mycoplasmas use single approach and do not confirm their findings by alternative methods. Consequently, the results of proteomic analysis should be validated by complementary techniques. In this study we utilized three complementary approaches (SWATH, MRM, 2D-DIGE) to assess response of Mycoplasma gallisepticum under heat stress on proteomic level and combined these findings with metabolic response and the results of transcriptional profiling. We divide response into two modes - one is directly related to heat stress and other is triggered during heat stress, but not directly relevant to it. The latter includes accumulation of ATP and shedding of antigens. Both of these phenomena may be relevant to evasion of host's immune system and dissemination during mycoplasmosis in vivo.


Subject(s)
Heat-Shock Response , Mycoplasma Infections/microbiology , Mycoplasma gallisepticum/physiology , Adenosine Triphosphate/metabolism , Computational Biology/methods , Hemagglutination Tests , Metabolome , Metabolomics , Temperature , Terpenes/metabolism
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