Proteomic landscape of seminal plasma associated with dairy bull fertility.

Male fertility is the ability of sperm to fertilize the egg and sustain embryo development. Several factors determine the fertilizing capacity of mammalian sperm, including those intrinsic to sperm and components of the seminal plasma. The present study analyzed the seminal fluid proteome of Bos taurus and potential associations between proteins and fertility scores. Mass spectrometry coupled with nano HPLC allowed the identification of 1,159 proteins in the dairy bull seminal plasma. There were 50 and 29 seminal proteins more abundant in high (HF) low fertility (LF) bulls, respectively. Based on multivariate analysis, C-type natriuretic peptide, TIMP-2, BSP5 and sulfhydryl oxidase indicated relationship with HF bulls. Clusterin, tissue factor pathway inhibitor 2, galectin-3-binding protein and 5'-nucleotidase were associated with LF bulls. Abundance of NAD(P)(+)-arginine ADP-ribosyltransferase, prosaposin and transmembrane protein 2 proteins had the highest positive correlations with fertility ranking. Quantities of vitamin D-binding protein, nucleotide exchange factor SIL1 and galectin-3-binding protein showed the highest negative correlations with fertility ranking. A fertility ranking score was calculated and the relationship with these proteins was significant (Spearman's rho = 0.94). The present findings represent a major and novel contribution to the study of bovine seminal proteins. Indicators of fertility can be used to improve reproductive biotechnologies.

Trypanosoma cruzi mitochondrial tryparedoxin peroxidase is located throughout the cell and its pull down provides one step towards the understanding of its mechanism of action.

Trypanosoma cruzi depends on the effectiveness of redox metabolism to survive and ensure infection in the host. Homeostasis of redox metabolism in T. cruzi is achieved by the actions of several proteins that differ in many aspects from host proteins. Although extensive research has been performed examining hydroperoxide cytosolic antioxidant defense centered on trypanothione, the mechanisms of mitochondrial antioxidant defense are not yet known. The aim of this study was to elucidate the partners of TcMPx antioxidant pathway and to determine the influence of the cellular context (physiological versus oxidative stress). Through co-precipitation coupled with a mass spectrometry approach, a variety of proteins were detected under physiological and oxidative stress conditions. Interestingly, functional category analysis of the proteins identified under physiological conditions showed that they were involved in the stress response, oxidoreduction, thiol transfer, and metabolic processes; this profile is distinct under oxidative stress conditions likely due to structural alterations. Our findings help to elucidate the reactions involving TcMPx and most importantly also reveal that this protein is present throughout the cell and that its interaction partners change following oxidative stress exposure. The involvement and significance of the proteins found to interact with TcMPx and other possible functions for this protein are discussed widening our knowledge regarding T. cruzi mitochondrial antioxidant defenses.

A C-type lectin from Bothrops jararacussu venom can adhere to extracellular matrix proteins and induce the rolling of leukocytes

Purification of a lectin from Bothrops jararacussu venom (BjcuL) was carried out using agarose-D-galactose affinity gel. MALDI-TOF gave a major signal at m/z 32028, suggesting the presence of a dimmer composed of two identical subunits. Divalent cations were required for the lectin activity, as complete absence of such ions reduced hemagglutination. BjcuL was more effective at neutral pH and showed total loss of activity at pH values below 4.0 and above 9.0. Its agglutinating activity remained stable at 25°C until 60min, but increased when at 35°C for at least 15min. Adhesion assays to extracellular matrix (ECM) glycoproteins showed that the biotinylated lectin (0.039-5.0µg/100µl) was capable of binding to fibronectin and vitronectin in a dose-dependent manner. The binding was partially inhibited in the presence of D-galactose. BjcuL (1.25-10µg/30µl) potential was investigated for leukocyte rolling and adhesion to endothelial cells in living microvessels using intravital microscopy, which showed that it induced a dose-dependent increase in rolling and adherence of leukocytes, acting directly on endothelial cells of postcapillary venules. The specific association between lectins and their ligands, either on the cell surface or on the ECM, is related to a variety of biological processes. The complementary characterization of BjcuL, shown here, is useful to further understand the venom effects and as a background for future investigation for therapeutic strategies.

Correlation between fertility and levels of protein, sugar and free amino acids in seminal plasma of Nelore bulls

Estimaram-se as correlações entre fertilidade e concentrações de proteínas, açúcares redutores e 17 tipos de aminoácidos livres do plasma seminal de 19 touros Nelore, adultos, doadores de sêmen de alta qualidade segundo as análises físicas e morfológicas. O escore de fertilidade foi estimado de acordo com a porcentagem de gestação das fêmeas cobertas em três estações de monta consecutivas, por inseminação artificial ou monta natural. Foi observada alta variabilidade na concentração das moléculas analisadas nas amostras. O coeficiente de correlação estimado entre concentrações de proteína e de açúcares redutores foi de 0,90. Individualmente, nenhum dos componentes quantificados apresentou correlação significativa com a fertilidade dos touros.

Correlation between fertility and levels of protein, sugar and free amino acids in seminal plasma of Nelore bulls / Correlação entre fertilidade e níveis de proteína, açúcar e aminoácidos livres no plasma seminal de touros Nelore

Correlations between fertility and the concentrations of proteins, reducing sugars and 17 types of free amino acids in the seminal plasma of Nelore bulls were estimated. The samples were collected from 19 bulls that had high quality semen, according to physical and morphological analyses. The bulls had their fertility scores estimated based on the pregnancy of the females mated to them during three consecutive breeding seasons, either by artificial insemination or by natural breeding. A large variability in the concentrations of the molecules was observed among the samples and a significant correlation coefficient (r= 0.90) between levels of proteins and reducing sugars was estimated. None of the components showed a significant correlation with fertility.(AU)

Estimaram-se as correlações entre fertilidade e concentrações de proteínas, açúcares redutores e 17 tipos de aminoácidos livres do plasma seminal de 19 touros Nelore, adultos, doadores de sêmen de alta qualidade segundo as análises físicas e morfológicas. O escore de fertilidade foi estimado de acordo com a porcentagem de gestação das fêmeas cobertas em três estações de monta consecutivas, por inseminação artificial ou monta natural. Foi observada alta variabilidade na concentração das moléculas analisadas nas amostras. O coeficiente de correlação estimado entre concentrações de proteína e de açúcares redutores foi de 0,90. Individualmente, nenhum dos componentes quantificados apresentou correlação significativa com a fertilidade dos touros.(AU)

Partial sequence and toxic effects of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera

A neurotoxic peptide, granulitoxin (GRX), was isolated from the sea anemone Bunodosoma granulifera. The N-terminal amino acid sequence of GRX is AKTGILDSDGPTVAGNSLSGT and its molecular mass is 4958 Da by electrospray mass spectrometry. This sequence presents a partial degree of homology with other toxins from sea anemones such as Bunodosoma caissarum, Anthopleura fuscoviridis and Anemonia sulcata. However, important differences were found: the first six amino acids of the sequence are different, Arg-14 was replaced by Ala and no cysteine residues were present in the partial sequence, while two cysteine residues were present in the first 21 amino acids of other toxins described above. Purified GRX injected ip (800 µg/kg) into mice produced severe neurotoxic effects such as circular movements, aggressive behavior, dyspnea, tonic-clonic convulsion and death. The 2-h LD50 of GRX was 400 ñ 83 µg/kg