Activity-based protein profiling of glycosidases in the vaginal microbiome
Ontology highlight
ABSTRACT:
INSTRUMENT(S): LTQ Orbitrap, Orbitrap Eclipse
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Cervicovaginal Fluid
DISEASE(S): Disease Free
SUBMITTER: 
Christopher Whidbey
LAB HEAD: Christopher Whidbey
PROVIDER: PXD042917 | Pride | 2023-08-04
REPOSITORIES: Pride
Similar Datasets
Project description:Cervical-vaginal fluid (CVF) covers the lower part of the female reproductive system and functions in the homeostasis and immunity of the surrounding tissues. The proteome of this proximal fluid has mainly been studied in pregnant women, whereas the CVF proteome of non-pregnant women has not been analyzed in great detail. The CVF peptidome has not been reported to date. In the current study, we separated pooled CVF samples from healthy non-pregnant women into proteomic and peptidomic fractions, followed by mass spectrometry analysis. In total, we identified 1,087 unique proteins in CVF, of which 801 proteins were not previously identified. The presence of the tissue specific proteins oviductal glycoprotein-1 (OVGP-1) and tubulin polymerization-promoting protein family member 3 (TPPP3) in CVF strongly suggests that the tissues of the upper female reproductive tract contribute to the protein composition of CVF. The tremendous catalytic potential of CVF was highlighted by the identification of 85 proteases. The majority of identified proteases belong to the serine protease catalytic type. Over 1,000 endogenous peptides were detected in the CVF peptidome, and 39 peptides are predicted to have antimicrobial activity. The detailed proteomic and peptidomic analysis of CVF will further aid in the delineation of physiological and pathobiological pathways related to reproduction, immunity and host defense, and assist in developing new biomarkers for malignant and other diseases of the female reproductive tract.
2017-09-27 | PXD004363 | Pride
Project description:Cervical-vaginal fluid (CVF) covers the lower part of the female reproductive system and functions in the homeostasis and immunity of the surrounding tissues. The proteome of this proximal fluid has mainly been studied in pregnant women, whereas the CVF proteome of non-pregnant women has not been analyzed in great detail. The CVF peptidome has not been reported to date. In the current study, we separated pooled CVF samples from healthy non-pregnant women into proteomic and peptidomic fractions, followed by mass spectrometry analysis. In total, we identified 1,087 unique proteins in CVF, of which 801 proteins were not previously identified in CVF. The presence of the tissue specific proteins oviductal glycoprotein-1 (OVGP-1) and tubulin polymerization-promoting protein family member 3 (TPPP3) in CVF strongly suggests that the tissues of the upper female reproductive tract contribute to the protein composition of CVF. The tremendous catalytic potential of CVF was highlighted by the identification of 85 proteases. The majority of identified proteases belonged to the serine protease catalytic type. Over 1,000 endogenous peptides were detected in the CVF peptidome, and 39 peptides are predicted to have antimicrobial activity. The detailed proteomic and peptidomic analysis of CVF will further aid in the delineation of physiological and pathobiological pathways related to reproduction, immunity and host defense, and assist in developing new biomarkers for malignant and other diseases of the female reproductive tract.
2017-09-27 | PXD004450 | Pride
Project description:Preterm birth (PTB) is one of major causes of perinatal mortality and neonatal morbidity, but knowledge of its complex etiology is still limited. Here we present cervicovaginal fluid (CVF) protein profiles of pregnant women who subsequently delivered at spontaneous preterm or term, aiming to identify differentially expressed CVF proteins in PTB and term birth. The CVF proteome of women who sequentially delivered at preterm and term was analyzed using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with two-dimensional nanoflow liquid chromatography-tandem mass spectrometry (2D-nLC-MS/MS). We compared the CVF proteome of PTB (n=5) and control subjects (term birth, n=7) using pooled control CVF (term birth, n=20) as spike-in standard. We identified 1294 CVF proteins, of which 605 were newly identified proteins. Of 990 proteins quantified in both PTB and term birth, 52 proteins were significantly up/down-regulated in PTB compared to term birth. The differentially expressed proteins were functionally associated to immune response, endopeptidase inhibitors and structural constituent of cytoskeleton. Taken together, our study provide quantitative CVF proteome profiles of pregnant women who ultimately delivered at preterm and term. These promising results could help to improve the understanding of PTB etiology and to discover biomarkers for asymptomatic PTB.
2021-09-09 | PXD021401 | Pride
Project description:Monocytes and macrophages are essential cells in the early response in their capacity as ubiquitous phagocytic cells. They phagocytose microorganisms or damaged cells and sense pathogen/damage-associated molecular patterns (PAMPs/DAMPs) through innate receptors such as Toll-like receptors (TLRs). We investigated a phenomenon where co-signaling from TLR2 and TLR8 in human primary monocytes provides a distinct immune activation profile compared to signaling from either TLR alone. Comparison of gene signatures induced by either stimulus alone or together, show that co-signaling result in downstream differences in regulation of signaling and gene transcription. To investigate the interaction of TLR2 and TLR8 stimulation primary human monocytes isolated from buffy coat were stimulated with TLR2/6 ligand FSL-1 and/or TLR7/8 ligand CL075 and the gene expression monitored at 1,2 and 3h post stimulation.
2018-03-09 | E-MTAB-6222 | biostudies-arrayexpress
Project description:Pneumocystis is a pathogen of immunocompromised hosts but can also infect healthy hosts, in whom infection is rapidly controlled and cleared. To better understand the immune mechanisms contributing to clearance of infection, microarray methods were used to examine differential gene expression in the lungs of C57BL/6 and CD40 ligand knock-out (CD40L-KO) mice over time following exposure to Pneumocystis. Immuncompetent C57BL/6 mice, which control and clear infection efficiently, showed a robust response to infection characterized by the upregulation of 349 primarily immune-response associated genes. Temporal changes in the expression of these genes suggested that there was an early (week 2) primarily innate response, that waned without controlling infection; this were followed by primarily adaptive immune responses that peaked at week 5 and successfully cleared the infection. In conjunction with the latter, there was an increased expression of B cell associated (immunoglobulin) genes at week 6 that persisted through 11 weeks. In contrast, CD40L-KO mice, which are highly susceptible to developing severe Pneumocystis pneumonia, showed essentially no upregulation of immune-response associated genes at days 35 to 75. Immunohistochemical staining supported these observations by demonstrating an increase in CD4+, CD68+, and CD19+ cells in C57BL/6 but not CD40L-KO mice. Thus, the healthy host demonstrates a robust biphasic response to infection by Pneumocystis; CD40 ligand is an essential upstream regulator of the adaptive immune responses that efficiently control infection and prevent development of progressive pneumonia. Keywords: Time course response Pneumocystis murina infection wild type versus CD40L-KO mice In experiment 1, C57BL/6 and CD40L-KO mice were co-housed in 2 cages; one cage was exposed to a P. murina-infected seeder, while the second was unexposed. Mice (3 per group) were sacrificed at day 32. Because the initial study demonstrated very little change in gene expression in CD40L-KO mice, only C57BL/6 mice were used for experiments 2 and 3. In Experiment 2, three cages (10 animals/cage) were set up: 2 cages were exposed to P. murina, and one was unexposed. Animals (5 per cage) were sacrificed at days 34 and 41. In Experiment 3, which focused on gene expression early after exposure to P. murina, five cages (9-10 animals/cage) were set up, of which 3 were exposed and 2 were unexposed. Animals (1-3 per cage) were sacrificed at days 7, 14 and 21 of exposure. Experiment 4 was designed to verify the initial results in CD40L-KO mice, and included 5 unexposed controls and 4 mice exposed for 35 days.
2008-12-22 | E-GEOD-11005 | biostudies-arrayexpress
Project description:In order to functionally characterise the LATERAL ORGAN BOUNDARIES DOMAIN (LBD) 37/38/39 proteins during ABA response we performed microarray analysis of Col-0 and lbd37 lb38 lbd39 treated with Mock or ABA.
2023-01-09 | E-MTAB-8688 | biostudies-arrayexpress
Project description:Arabidopsis wild type and atglr3.3 mutant seedlings were cultured on 0.7% (w/v) agar medium containing half strength of MS basic salt with 1% sucrose (w/v), 5mM MES, pH 5.7. 10-day-old seedlings were equilibrated in the CK buffer for 24 hours. Then, these seedlings were either treated with the CK buffer or the CK buffer containing 100 ?M GSH and 100 ?M Glu for 1 hour, respectively.
2013-09-30 | E-MEXP-3885 | biostudies-arrayexpress
Project description:Dysregulation of microRNAs (miRNAs) plays a critical role in the development of intervertebral disc degeneration (IDD). In this study, we present evidence from in vitro and in vivo research to elucidate the mechanism underlying the role of miR-760 in IDD. miRNA microarray and quantitative reverse transcription-polymerase chain reaction were used to determine the miRNA profiles in patients with IDD. Functional analysis was performed to evaluate the role of miR-760 in the pathogenesis of IDD. Luciferase reporter and western blotting assays were used to confirm the miRNA targets. The expression of miR-760 was significantly decreased in degenerative nucleus pulposus (NP) cells and negatively correlated with disc degeneration grade. Functional assays demonstrated that miR-760 delivery significantly increased NP cell proliferation and promoted the expression of collagen II and aggrecan. Moreover, MyD88 was identified as a target gene of miR-760. miR-760 effectively suppressed MyD88 expression by interacting with the 3′-untranslated region, which was abolished by miR-760 binding site mutations. An in vivo experiment using an IDD mouse model showed that the upregulation of miR-760 could effectively suspend IDD. Therefore, miR-760 was found to play an important role in IDD and can be used as a promising therapeutic target for the treatment of patients with IDD.
2022-03-17 | E-MTAB-11554 | biostudies-arrayexpress
Project description:There were two genotypes:,(1) Columbia-0, wild-type (C),(2) pmr4-1 mutant (P), callose synthase deficient mutant (Vogel and Somerville (2000) Proc. Natl. Acad. Sci., USA 97: 1897).,There were two treatments:,(1) uninoculated (U),(2) 3 days after inoculation with the powdery mildew pathogen, Erysiphe cichoracearum, race UCSC (I).,There were four biological replicates, labeled 1, 2, 3 or 4.,Examples of the sample labels are:,CU1 = Columbia-0, uninoculated, replicate 1,CI2 = Columbia-0, 3 days after inoculation with powdery mildew, replicate 2,PU3 = pmr4-1, uninoculated, replicate 3,PI4 = pmr4-1, 3 days after inoculation with powdery mildew, replicate 4.,In total, there were 16 Affymetrix ATH1 GeneChips (2 genotypes x 2 treatments x 4 biological replicates).
2007-07-13 | E-GEOD-431 | biostudies-arrayexpress
Project description:Array experiment: Animals were divided into a control group (naive), which did not undergo any experimental manipulation (n=20) and two experimental groups, one receiving an intra-orbital nerve transection (IONT, n=60) and the other an intra-orbital nerve crush (IONC, n=60) Sterile precautions were maintained for all surgical procedures. The left ON was intraorbitally axotomized according to procedures that are standard in our laboratory. (REF) Briefly, an incision was made in the superior orbital rim, the superoexternal orbital contents were dissected, and the superior and external rectus muscles were removed. When performing IONT, the dura mater was opened longitudinally to spare the blood supply, and the optic nerve was transected 0.5 mm from the optic disc. To perform IONC the optic nerve was crushed 3 mm from the optic disc during 10 seconds using a pair of watchmakers forceps (Vidal-Sanz et al., 1988, 1991; Villegas-Perez et al., 1993). Before and after the procedure, the eye fundus was observed through the operating microscope to assess the integrity of the retinal blood flow. Tissue processing and RNA extraction: Animals were kept the appropriate time post injury (12h, 24h, 48h, 3d 7d and 15d, n=8-12 per time point) and then sacrificed by an intraperitoneal overdose of sodium pentobarbital. Left retinas were fresh dissected and immediately frozen in liquid nitrogen. Four retinas from each time point, animal group and biological replica were pooled and RNA was extracted using Trizol (Invitrogen). RNA was further cleaned through RNAeasy mini kit columns (Quiagen). The RNA integrity was checked using a Bioanalyzer (Agilent Technologies, USA) and concentrations were determined using a NanoDrop (NanoDrop Technologies, Wilmington, DE, USA).
2008-06-14 | E-MEXP-1387 | biostudies-arrayexpress