Workflow for metaproteome characterization in bronchoalveolar lavage fluid (BALF)
Ontology highlight
ABSTRACT:
INSTRUMENT(S): LTQ Orbitrap Velos
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Bronchoalveolar Lavage
DISEASE(S): Respiratory Failure
SUBMITTER: 
Kevin Viken
LAB HEAD: Maneesh Bhargava
PROVIDER: PXD008273 | Pride | 2019-06-24
REPOSITORIES: Pride
Publications
BAL Fluid Metaproteome in Acute Respiratory Failure.
Jagtap Pratik D PD Viken Kevin J KJ Johnson James J McGowan Thomas T Pendleton Kathryn M KM Griffin Timothy J TJ Hunter Ryan C RC Rudney Joel D JD Bhargava Maneesh M
American journal of respiratory cell and molecular biology 20181101 5
Similar Datasets
Project description:The human cervical-vaginal area contains proteins derived from microorganisms that may prevent or predispose women to gynecological conditions. The liquid Pap test fixative is an unexplored resource for analysis of microbial communities and the microbe-host interaction. Previously, we showed that the residual cell-free fixative from discarded Pap tests of healthy women could be used for mass spectrometry (MS) based proteomic identification of cervical-vaginal proteins. In this study, we reprocessed these MS raw data files for metaproteomic analysis to characterize the microbial community composition and function of microbial proteins in the cervical-vaginal region. This was accomplished by developing a customized protein sequence database encompassing microbes likely present in the vagina. High-mass accuracy data were searched against the protein FASTA database using a two-step search method within the Galaxy for proteomics platform. Data was analyzed by MEGAN6 (MetaGenomeAnalyzer) for phylogenetic and functional characterization. We identified over 300 unique peptides from a variety of bacterial phyla and Candida. Peptides corresponding to proteins involved in carbohydrate metabolism, oxidation-reduction, and transport were identified. By identifying microbial peptides in Pap test supernatants it may be possible to acquire a functional signature of these microbes, as well as detect specific proteins associated with cervical health and disease.
2018-07-23 | PXD009596 | Pride
Project description:The lesions of enamel caries can be considered as the outcome of dysbiotic changes in the biofilm community of supragingival dental plaque. Demineralization occurs as the cumulative outcome of repeated shifts towards a less diverse microbiota that produces and tolerates a low pH environment in tooth sites that are sheltered from protective factors in host saliva. Although the etiology of caries is multifactorial, frequent consumption of foods rich in fermentable carbohydrates, notably sucrose, appears to be one of the major factors driving the microbiota in the direction of dysbiosis, particularly in the case of otherwise healthy children with normal salivary flow. Streptococcus mutans and closely related species (such as Streptococcus sobrinus) have long been considered to play a primary etiological role in dental caries. S. mutans responds to sucrose by producing large quantities of lactic acid. It is very tolerant of low pH, and produces an insoluble extracellular polysaccharide that may sequester acid at tooth surfaces. The mechanisms behind those putative virulence factors have been intensively studied in monoculture, and recently in simple multi-species consortia. Much less is known of other species that may also contribute to or protect against dysbiosis driven by dietary carbohydrates. Some strains of “non-mutans” streptococci produce and tolerate acid at levels comparable to S. mutans, while others show increased ariginolytic capabilities, which may act to raise pH within the biofilm matrix. S. mutans tends to be a minority species even in caries-active children, and carious lesions likewise can occur in children with no detectable S. mutans. 16S rDNA-based metagenomic comparisons of caries-active and caries-free subjects have detected associations between caries and a variety of oral species, including not only non-mutans streptococci, but also members of other genera, such as Scardovia and Bifidobacterium. Caries associations have not been consistent between studies. Moreover, different taxonomic clusters have been defined as subgroups within the same study. This raises an important point. Although caries-associated communities are typically less diverse than healthy supragingival plaque overall, those dysbiotic communities still display considerable taxonomic diversity between affected individuals. That in turn raises the question of whether it is desirable to define biomarkers of dysbiosis that are less dependent on taxonomy. The Human Microbiome project generated comprehensive metagenomic data for a wide variety of body sites in healthy subjects, including supragingival plaque. Although most of that data was based on 16S rDNA sequencing, shotgun metagenomics was also used to catalog the functional potential of all microbial genes within a smaller subset of subjects. One of the key findings was that healthy sites from different people were broadly similar with respect to their functional profiles, even though there was extensive individual variation in their taxonomic profiles. It is possible that the “conservation of function” concept may also extend to dysbiotic communities. This would explain why microbial communities associated with caries still show considerable taxonomic variation. In that case, differential patterns of community-wide gene and/or protein expression might provide a more accurate indicator of dysbiosis than can be achieved by counting caries-associated species. Metatranscriptomic or metaproteomic approaches can be used to provide information on function. A recent metatranscriptomic comparison of subgingival plaque from healthy and periodontally diseased sites in three subjects has provided data that support the “conservation of function” concept. They observed that taxonomically diverse diseased sites shared conserved gene expression profiles [20]. By the same token, a recent metaproteomic comparison of gut microbiotas from healthy controls to Crohn’s disease patients found that major shifts in protein expression by function did not always correlate with changes in taxon relative abundance [21]. In this metaproteomic study, we found that sucrose–induced changes in protein expression patterns for pathways involving glycolysis, lactate production, aciduricity and ammonia/glutamate metabolism were likewise conserved in taxonomically diverse dysbiotic oral microcosm biofilm communities.
2016-01-04 | PXD003151 | Pride
Project description:Acute Respiratory Distress Syndrome (ARDS) continues to have a high mortality. The objective of this study is to understand the differences in disease biology between survivors and non-survivors by characterizing BALF protein expression profiles in individual ARDS subjects.
2018-10-24 | PXD002672 | Pride
Project description:The goal of this study was to apply protein expression profiling tools in cases with lung injury following HSCT or cellular therapy infusion to identify the proteins and biological processes that differentiate IPS from infectious lung injury in HSCT recipients. We performed comprehensive label-based quantitative protein profiling of BALF in patients undergoing HSCT
2018-10-24 | PXD002437 | Pride
Project description:Up to date, most metaproteomic studies of the gut microbiota describe the use of stool sample pretreatment methods to enrich for microbial components. However, a specific investigation aimed at assessing if, how and to what extent this may impact on the final taxonomic and functional results is still lacking. Here, stool replicates were either pretreated by differential centrifugation (DC) or not centrifuged (NC). Protein extracts were then processed by filter-aided sample preparation, single-run LC and high-resolution MS, and the metaproteomic data were compared by spectral counting. DC led to a higher number of identifications, a significantly richer microbial diversity, as well as to reduced information on the non-microbial components (host and food) when compared to NC. Nevertheless, dramatic differences in the relative abundance of many gut microbial taxa were also observed, including a significant change in the Firmicutes/Bacteroidetes ratio. Furthermore, several microbial functional categories, including cell surface enzymes, membrane-associate proteins, and flagella, were significant reduced after DC. In conclusion, this work underlines that a critical evaluation is needed when selecting the appropriate stool sample processing protocol in the context of a metaproteomic study, depending on the specific target to which the research is aimed.
2015-02-25 | PXD001573 | Pride
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
2015-02-25 | PXD001361 | Pride
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
2015-02-25 | PXD001357 | Pride
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
2015-02-26 | PXD001362 | Pride
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
2015-02-26 | PXD001359 | Pride
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
2015-02-25 | PXD001360 | Pride