RESUMO
Mexico has the highest diversity of snake species in the world, following Australia when considering just venomous snakes. Specifically, in Sonora, the second largest state in the country, more than 15 highly venomous species occur, including the northern black-tailed rattlesnake (Crotalus molossus). This specie's venom has not been as thoroughly researched in contrast with other Mexican vipers, nevertheless some studies report its biological activity and even pharmacological potential with antibacterial and cytotoxic activity. In this study we identified the main protein components from a pool of C. molossus venom through a gel-free proteomics approach, reporting â¼140 proteins belonging to the SVMP (38.76%), PLA2 (28.75%), CTL (11.93%), SVSP (6.03%) and LAAO (5.67%) toxin families. To study its biological activities, we evaluated its hemolytic, antibacterial, and cytotoxic activity in red blood cells, Gram positive and negative bacteria and a luminal A breast carcinoma cell line (T47D), respectively, in vitro. We report that concentrations <100 µg/mL are potentially not hemolytic and reduced the bacteria viability of E. coli and S. aureus with an IC50 of 10.27 and 11.51 µg/mL, respectively. Finally, we determined the C. molossus venom as cytotoxic against the T47D breast carcinoma cell line, with an IC50 of 1.55 µg/mL. We suggest that the evaluated cytotoxicity was due to a high abundance of SVMPs and PLA2s, since it's been reported that they affect the extracellular matrix and membrane permeation. This may provide a useful tool for pharmaceutical screening in the future.
RESUMO
This data article contains data related to the research article titled Proteomic analysis of chloroplast biogenesis (clb) mutants uncovers novel proteins potentially involved in the development of Arabidopsis thaliana chloroplasts (de Luna-Valdez et al., 2014) [1]. This research article describes the 2-D PAGE-based proteomic analysis of wild-type and four mutant lines (cla1-1, clb2, clb5 and clb19) affected in the development of Arabidopsis thaliana chloroplasts. The report concludes with the discovery of three proteins potentially involved in chloroplast biogenesis. The information presented here represent the tables and figures that detail the processing of the raw data obtained from the image analysis of the 2-D PAGE gels.
RESUMO
Plant cells outstand for their ability to generate biomass from inorganic sources, this phenomenon takes place within the chloroplasts. The enzymatic machinery and developmental processes of chloroplasts have been subject of research for several decades, and this has resulted in the identification of a plethora of proteins that are essential for their development and function. Mutant lines for the genes that code for those proteins, often display pigment-accumulation defects (e.g., albino phenotypes). Here, we present a comparative proteomic analysis of four chloroplast-biogenesis affected mutants (cla1-1, clb2, clb5, clb19) aiming to identify novel proteins involved in the regulation of chloroplast development in Arabidopsis thaliana. We performed 2D-PAGE separation of the protein samples. These samples were then analyzed by computational processing of gel images in order to select protein spots with abundance shifts of at least twofold, statistically significant according to Student's t-test (P<0.01). These spots were subjected to MALDI-TOF mass-spectrometry for protein identification. This process resulted in the discovery of three novel proteins potentially involved in the development of A. thaliana chloroplasts, as their associated mutant lines segregate pigment-deficient plants with abnormal chloroplasts, and altered mRNA accumulation of chloroplast-development marker genes. BIOLOGICAL SIGNIFICANCE: This report highlights the potential of using a comparative proteomics strategy for the study of biological processes. Particularly, we compared the proteomes of wild-type seedlings and four mutant lines of A. thaliana affected in chloroplast biogenesis. From this proteomic analysis it was possible to detect common mechanisms in the mutants to respond to stress and cope with heterotrophy. Notably, it was possible to identify three novel proteins potentially involved in the development or functioning of chloroplasts, also it was demonstrated that plants annotated to carry T-DNA insertions in the cognate genes display pigment-deficient phenotypes, aberrant and underdeveloped chloroplasts, as well as altered mRNA accumulation of chloroplast biogenesis marker genes.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Cloroplastos/metabolismo , Mutação , Proteômica , Arabidopsis/genética , Cloroplastos/patologia , Análise por Conglomerados , DNA Bacteriano/genética , Eletroforese em Gel Bidimensional , Perfilação da Expressão Gênica , Heterozigoto , Pigmentação , Proteoma , RNA Mensageiro/metabolismo , Plântula/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Worldwide, cervical and uterine cancers are the most deadly cancers in women, with high prevalences, especially in developing countries. The Human Protein Atlas (HPA) portal was explored for proteins expressed in a tissue- or cervix and uterine cancer-specific manner. The group of proteins differentially expressed and with enhanced expression in the glandular and surface epithelial (squamous) cells retrieved from HPA were further explored using the Protein Information and Knowledge Extractor (PIKE) portal to compile biological information that is found in different databases, and repositories on the Internet. Thus, the lists of candidate proteins found in HPA, and PIKE portals may be used as a starting point for the discovery and validation of biomarkers for cervix and uterine cancer employing proteomics approaches as described in the present article (AU)
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