The present study was undertaken to identify potentially immunoreactive proteins of the muscle larvae (ML) and adult stage (Ad) of the nematode Trichinella spiralis Owen, 1835. To identify immunoreactive proteins that are specifically recognised by anti-Trichinella antibodies, ML and Ad crude extracts and their excretory-secretory (E-S) products were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot with serum samples from pigs experimentally infected with T. spiralis. A total of 18 bands were selected for final identification by liquid chromatography-tandem mass spectrometry. To further understand the functions of the proteins identified in this study, gene ontology terms were applied. Results showed that the specific antibodies against T. spiralis reacted with protein bands matching heat shock proteins, aminopeptidase, enolase, isocitrate dehydrogenase NADP-dependent, tropomyosin, P49 antigen, serine proteinase, secreted 5'-nucleotidase, antigen targeted by protective antibodies, 53 kDa E-S antigen, putative trypsin and paramyosin. Three proteins common for both adult stage and muscle larvae, including heat shock proteins, enolase and 5'-nucleotidase, might play important role during T. spiralis infection. These proteins are presumably presented to the host immune system and may induce humoral immune response. Thus, these proteins may be potential antigens for early diagnosis and the development of a vaccine against the parasite., Justyna Bien, Wladyslaw Cabaj, Bozena Moskwa., and Obsahuje bibliografii
To determine the relationship between protein expression and insect diapause, a proteomic approach was used to investigate the proteins extracted from larvae of the wheat blossom midge Sitodiplosis mosellana Gehin at different developmental stages, including pre-diapause, over-summering diapause, over-wintering diapause and post-diapause. Using 2-DE gels stained with coomassie brilliant blue, about 300 protein spots were detected in the extracts of pre-diapause larvae and 275 for those in each of the other stages. There were 91, 92 and 95 protein spots that showed more than a 2-fold change in abundance in the over-summering diapause, over-wintering diapause and post-diapause stages compared with pre-diapause. Eight protein spots, which showed the greatest difference in the larvae at different stages of diapause, were analyzed using Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Seven of them were successfully identified from their peptide mass fingerprints using the NCBInr database. They were proopiomelanocortin, NADH dehydrogenase subunit 1 and F10F2.5, which were up-regulated or unique to pre-diapause larvae, IKK interacting protein isoform 2 up-regulated in diapause and post-diapause larvae, GA10647-PA unique in over-wintering diapause larvae, purple CG16784-PB isoform B and B0228.6 up-regulated in over-summering and over-wintering diapause larvae. The potential functions of these proteins during wheat blossom midge diapause are discussed.
Saliva contains possible biomarkers that are associated with dental caries. The present study aimed to analyse differences in the abundance of proteins in the saliva between caries-positive (CP; N = 15) and caries-free (CF; N = 12) males and to compare differences in the abundance of proteins between two saliva sample fractions (supernatant and pellet). We found 14 differently significantly expressed proteins in the CF group when comparing the supernatant fractions of the CP and CF groups, and three proteins in the pellet fractions had significantly higher expression in the CP group. Our results indicate very specific protein compositions of the saliva in relation to dental caries resistance (the saliva of the CP group mainly contained pellet proteins and the saliva of the CF group mainly contained supernatant proteins). This was the first time that the saliva pellet fraction was analysed in relation to the dental caries status. We detected specific calcium-binding proteins that could have decalcified enamel in the saliva pellet of the CP group. We also observed significantly up-regulated immune proteins in the saliva supernatant of the CF group that could play an important role in the caries prevention. The particular protein compositions of the saliva pellet and supernatant in the groups with different susceptibilities to tooth decay is a promising finding for future research.
Proteinuria is often used as a surrogate marker in monitoring and predicting outcome in patients with chronic kidney diseases, but it is non-specific. IgAN belongs to the most common primary glomerulonephritis worldwide with serious prognosis. The main aim of this work was to assess differences in urine proteins in patients with IgA nephropathy and to identify abnormal proteins as potential biomarkers of IgA nephropathy or the renal disease. In our pilot project, we selected 20 patients and compared them with 20 healthy volunteers. Protein quantification was performed using iTRAQ (isobaric tag for relative and absolute quantitation) labeling method. The peptides were separated by the isoelectric focusing method (IEF) and nano-LC with C18 column and identified by mass spectrometry using MALDI-TOF/TOF MS. Proteins´ lists obtained from IEF-LC-MS-MS/MS analysis were combined and contained 201 proteins. It was found out that 113 proteins were common in both experiments. 30 urinary proteins were significantly up- or down-regulated in patients with IgA nephropathy. We characterized potential biomarkers such as alpha-1-antitrypsin, apolipoprotein A-I, CD44 antigen or kininogen. Potential biomarkers of IgAN should be validated in further studies., P. Prikryl, L. Vojtova, D. Maixnerova, M. Vokurka, M. Neprasova, T. Zima, V. Tesar., and Obsahuje bibliografii
Progeny of the flesh fly Sarcophaga bullata exposed to short day length show a maternal effect that prevents the expression of pupal diapause. Although ecological aspects of this effect are well studied, not enough is known about the molecular mechanisms underlying this maternal effect. In this study, two-dimensional electrophoresis was performed to detect differences of the abundance of certain proteins in the ovaries of this fly kept under long day and short day conditions for 2 days after eclosion. Eleven proteins that were abundant and showed significant changes were selected for mass spectrometric identification. Ovary proteins that increased in abundance under short-day conditions were similar to twinstar CG4254-PA, muscle protein 20-like protein, GA13413-PA, gene analogous to small peritrophins (Gasp CG10287-PA) and Ribosomal protein LP1 CG4087-PA. Ovary proteins that decreased in abundance under short-day conditions were similar to the ATP synthase beta subunit, fk506-binding protein and storage protein-binding protein. The 2-D proteome maps included 2 additional unknown proteins that were more abundant and 1 that was less abundant in the ovaries of 2-day old short-day females. Twinstar CG4254-PA, muscle protein 20-like protein and GA13413-PA harbour an actin-binding domain. That the 3 actin-binding proteins increase in abundance suggests that it is likely that an alteration in the actin cytoskeleton is involved in this maternal effect in the flesh fly., Aiqing Li ... [et al.]., and Obsahuje seznam literatury
Transcription factors exert their regulatory potential on RNA polymerase II machinery through a multiprotein complex called Mediator complex or Mediator. The Mediator complex integrates regulatory signals from cell regulatory cascades with the regulation by transcription factors. The Mediator complex consists of 25 subunits in Saccharomyces cerevisiae and 30 or more subunits in multicellular eukaryotes. Mediator subunit 28 (MED28), along with MED30, MED23, MED25 and MED26, belong to presumably evolutionarily new subunits that seem to be absent in unicellular eukaryotes and are likely to have evolved together with multicellularity and cell differentiation. Previously, we have shown that an originally uncharacterized predicted gene, F28F8.5, is the true MED28 orthologue in Caenorhabditis elegans (mdt-28) and showed that it is involved in a spectrum of developmental processes. Here, we studied the proteomic interactome of MDT-28 edited as GFP::MDT-28 using Crispr/Cas9 technology or MDT-28::GFP expressed from extrachromosomal arrays in transgenic C. elegans exploiting the GFPTRAP system and mass spectrometry. The results show that MDT-28 associates with the Head module subunits MDT-6, MDT-8, MDT-11, MDT-17, MDT20, MDT-22, and MDT-30 and the Middle module subunit MDT-14. The analyses also identified additional proteins as preferential MDT-28 interactants, including chromatin-organizing proteins, structural proteins and enzymes. The results provide evidence for MDT-28 engagement in the Mediator Head module and support the possibility of physical (direct or indirect) interaction of MDT-28 with additional proteins, reflecting the transcription-regulating potential of primarily structural and enzymatic proteins at the level of the Mediator complex.
Teeth have been a focus of interest for many centuries - due to medical problems with them. They are the hardest part of the human body and are composed of three mineralized parts - enamel, dentin and cementum, together with the soft pulp. However, saliva also has a signif icant impact on tooth quality. Proteomic research of human teeth is now accelerating, and it includes all parts of the tooth. Some methodological problems still need to be overcome in this research field - mainly connected with calcified tissues. This review will provide an overview of the current state of research with focus on the individual parts of the tooth and pellicle layer as well as saliva. These proteomic results can help not only stomatology in terms of early diagnosis, identifying risk factors, and systematic control., M. Jágr ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Vyšetření moči poskytuje důležité informace o patologických změnách probíhajících v ledvinách. Dosavadní vyšetření zaměřené obvykle jen na celkovou proteinurii, eventuálně albuminurii, zdaleka nevyužívá obrovské množství informací o fyziologických a patofyziologických procesech v ledvinách, které lze potenciálně získat proteomickou analýzou vzorku moči. Studium močového proteomu může vést k identifikaci nových markerů akutních i chronických onemocnění ledvin. Proteomická analýza dialyzátu získaného při hemodialýze může pomoci v nalezení nových uremických toxinů a markerů účinnosti očišťovacích metod. Studium proteomu jednotlivých renálních tkání (např. kůra vs dřeň) nebo buněčných populací, eventuálně buněčných kompartmentů (např. organel) může přispět k pochopení patogeneze renálních chorob a pochopení účinků farmakologické léčby. Další možností je využití cílené proteomiky (např. studium proteinů s určitými posttranslačními modifikacemi)., Examination of the urine provides us with the important information about the pathological changes affecting the kidney. Routine examination usually aimed only at total proteinuria or albuminuria does not exploit the huge amount of information about the physiological and pathophysiological processes in the kidney which can be potentially derived from the proteomic analysis of the sample of the urine. Study of the urinary proteome may lead to the identification of new markers of both acute and chronic kidney diseases. Proteomic analysis of the dialysate effluent obtained during hemodialysis may help to find new uremic toxins and markers of the effectivity of the blood purification methods. Study of the proteome of renal tissue (e. g. cortex vs. medulla), or cell populations, or cellular compartments (e. g. organelles) could contribute to the better understanding of the pathogenesis of the renal diseases and the effects of their pharmacological treatment. Another option is the use of the targeted proteomics (e. g. the study of proteins with defined posttranslational modifications)., Vladimír Tesař, Lucie Vojtová, Tomáš Zima, and Lit.: 35