Advances in the proteomic investigation of the cell secretome

Brown, Kristy J.; Formolo, Catherine A.; Seol, Haeri; Marathi, Ramya; Duguez, Stéphanie; An, Eunkyung; Pillai, Dinesh K.; Nazarian, Javad; Rood, Brian R.; Hathout, Yetrib

doi:10.1586/epr.12.21

Cited by 103 publications

(97 citation statements)

References 73 publications

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“…biomarker or therapeutic target discovery [15]. To do that, we chose one of the identified proteins, IL-33, and conducted a “proof-of-concept” experiment.…”

Section: Discussionmentioning

confidence: 99%

Global secretome characterization of A549 human alveolar epithelial carcinoma cells during Mycoplasma pneumoniae infection

Wang

et al. 2014

BMC Microbiol

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BackgroundMycoplasma pneumoniae (M. pneumoniae) is one of the major etiological agents for community-acquired pneumonia (CAP) in all age groups. The early host response to M. pneumoniae infection relies on the concerted release of proteins with various biological activities. However, no comprehensive analysis of the secretory proteins has been conducted to date regarding the host response upon M. pneumoniae infection.ResultsWe employed the liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based label-free quantitative proteomic technology to identify and characterize the members of the human alveolar epithelial carcinoma A549 cell secretome during M. pneumoniae infection. A total of 256 proteins were identified, with 113 being differentially expressed (>1.5-fold change), among which 9 were only expressed in control cells, 10 only in M. pneumoniae-treated cells, while 55 were up-regulated and 39 down-regulated by M. pneumoniae. The changed expression of some of the identified proteins was validated by RT-PCR and immunoblot analysis. Cellular localization analysis of the secretome data revealed 59.38% of the proteins were considered as “putative secretory proteins”. Functional analysis revealed that the proteins affected upon M. pneumoniae infection were mainly related to metabolic process, stress response, and immune response. We further examined the level of one up-regulated protein, IL-33, in clinical samples. The result showed that IL-33 levels were significantly higher in the plasma and bronchoalveolar lavage fluid (BALF) of M. pneumoniae pneumonia (MPP) patients.ConclusionsThe present study provided systematic information about the changes in the expression of secretory proteins during M. pneumoniae infection, which is useful for the discovery of specific biomarkers and targets for pharmacological intervention.

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“…biomarker or therapeutic target discovery [15]. To do that, we chose one of the identified proteins, IL-33, and conducted a “proof-of-concept” experiment.…”

Section: Discussionmentioning

confidence: 99%

Global secretome characterization of A549 human alveolar epithelial carcinoma cells during Mycoplasma pneumoniae infection

Wang

et al. 2014

BMC Microbiol

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“…While efforts are being made to improve secretome studies in vivo [3,20] several in vitro studies have generated valuable data that can be interrogated in vivo using human tissue biopsies and/or human body fluids [3]. However, one of the challenges in in vitro secretome studies is discriminating truly cell released proteins from proteins that are released due to cell death or leakage during culturing.…”

Section: Discussionmentioning

confidence: 99%

“…Typically the majority of secretome studies published to date are performed in vitro by culturing cells of interest first in serum supplemented medium to obtain sufficient number of cells then in serum free medium for a short period of time [3]. This allows collection of genuine cell secreted proteins while minimizing bovine serum interfering proteins.…”

Section: Introductionmentioning

confidence: 99%

The human secretome atlas initiative: Implications in health and disease conditions

Brown

Seol

Pillai

et al. 2013

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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Proteomic analysis of human body fluids is highly challenging, therefore many researchers are redirecting efforts towards secretome profiling. The goal is to define potential biomarkers and therapeutic targets in the secretome that can be traced back in accessible human body fluids. However, currently there is a lack of secretome profiles of normal human primary cells making it difficult to assess the biological meaning of current findings. In this study we sought to establish secretome profiles of human primary cells obtained from healthy donors with the goal of building a human secretome atlas. Such an atlas can be used as a reference for discovery of potential disease associated biomarkers and eventually novel therapeutic targets. As a preliminary study, secretome profiles were established for six different types of human primary cell cultures and checked for overlaps with the three major human body fluids including plasma, cerebrospinal fluid and urine. About 67% of the 1054 identified proteins in the secretome of these primary cells occurred in at least one body fluid. Furthermore, comparison of the secretome profiles of two human glioblastoma cell lines to this new human secretome atlas enabled unambiguous identification of potential brain tumor biomarkers. These biomarkers can be easily monitored in different body fluids using stable isotope labeled standard proteins. The long term goal of this study is to establish a comprehensive online human secretome atlas for future use as a reference for any disease related secretome study.

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“…Twenty-eight percent of the apical proteins were classified as cellular ( Figure E2A). Because recent publications show that numerous proteins previously not categorized as secreted are consistently present in in vivo and in vitro secretions (34,40), we compared our list of cellular proteins classified by current data bases with those published in The Human Secretome Atlas Initiative (34), resulting in 55% of the cellular proteins being reclassified as secreted and bringing the total percentage of secreted proteins to 81.7% ( Figure E2B). …”

Section: Comparison Of Secretomes From Normal Differentiated Hbe Primmentioning

confidence: 99%

Quantitative Proteomics Reveals an Altered Cystic FibrosisIn VitroBronchial Epithelial Secretome

Peters-Hall¹,

Brown

Pillai

et al. 2015

Am J Respir Cell Mol Biol

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Alterations in epithelial secretions and mucociliary clearance contribute to chronic bacterial infection in cystic fibrosis (CF) lung disease, but whether CF lungs are unchanged in the absence of infection remains controversial. A proteomic comparison of airway secretions from subjects with CF and control subjects shows alterations in key biological processes, including immune response and proteolytic activity, but it is unclear if these are due to mutant CF transmembrane conductance regulator (CFTR) and/or chronic infection. We hypothesized that the CF lung apical secretome is altered under constitutive conditions in the absence of inflammatory cells and pathogens. To test this, we performed quantitative proteomics of in vitro apical secretions from air-liquid interface cultures of three lifeextended CF (DF508/DF508) and three non-CF human bronchial epithelial cells after labeling of CF cells by stable isotope labeling with amino acids in cell culture. Mass spectrometry analysis identified and quantitated 666 proteins across samples, of which 70 exhibited differential enrichment or depletion in CF secretions (61.5-fold change; P , 0.05). The key molecular functions were innate immunity (24%), cytoskeleton/extracellular matrix organization (24%), and protease/antiprotease activity (17%). Oxidative proteins and classical complement pathway proteins that are altered in CF secretions in vivo were not altered in vitro. Specific differentially increased proteins-MUC5AC and MUC5B mucins, fibronectin, and matrix metalloproteinase-9-were validated by antibody-based assays. Overall, the in vitro CF secretome data are indicative of a constitutive airway epithelium with altered innate immunity, suggesting that downstream consequences of mutant CFTR set the stage for chronic inflammation and infection in CF airways.

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Advances in the proteomic investigation of the cell secretome

Cited by 103 publications

References 73 publications

Global secretome characterization of A549 human alveolar epithelial carcinoma cells during Mycoplasma pneumoniae infection

Global secretome characterization of A549 human alveolar epithelial carcinoma cells during Mycoplasma pneumoniae infection

The human secretome atlas initiative: Implications in health and disease conditions

Quantitative Proteomics Reveals an Altered Cystic FibrosisIn VitroBronchial Epithelial Secretome

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