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Plasma Membrane Profiling Reveals Upregulation of ABCA1 by Infected Macrophages Leading to Restriction of Mycobacterial Growth.

Long J, Basu Roy R, Zhang YJ, Antrobus R, Du Y, Smith DL, Weekes MP, Javid B - Front Microbiol (2016)

Bottom Line: The plasma membrane represents a critical interface between the internal and extracellular environments, and harbors multiple proteins key receptors and transporters that play important roles in restriction of intracellular infection.We quantified 559 PM proteins in BCG-infected THP-1 cells.One significantly upregulated cell-surface protein was the cholesterol transporter ABCA1.

View Article: PubMed Central - PubMed

Affiliation: Collaboration Innovation Centre for the Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University Beijing, China.

ABSTRACT
The plasma membrane represents a critical interface between the internal and extracellular environments, and harbors multiple proteins key receptors and transporters that play important roles in restriction of intracellular infection. We applied plasma membrane profiling, a technique that combines quantitative mass spectrometry with selective cell surface aminooxy-biotinylation, to Bacille Calmette-Guérin (BCG)-infected THP-1 macrophages. We quantified 559 PM proteins in BCG-infected THP-1 cells. One significantly upregulated cell-surface protein was the cholesterol transporter ABCA1. We showed that ABCA1 was upregulated on the macrophage cell-surface following infection with pathogenic mycobacteria and knockdown of ABCA1 resulted in increased mycobacterial survival within macrophages, suggesting that it may be a novel mycobacterial host-restriction factor.

No MeSH data available.


Related in: MedlinePlus

Gene ontology annotations of proteins identified by two or more peptides in each experiment. In Experiment A, peptides were fractionated by HpRp, and in Experiment B (label swap) unfractionated. Short GO terms include the annotations ‘membrane’, ‘integral to membrane’, ‘intrinsic to membrane’, ‘cell part’, but have no information as to subcellular localization.
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Figure 1: Gene ontology annotations of proteins identified by two or more peptides in each experiment. In Experiment A, peptides were fractionated by HpRp, and in Experiment B (label swap) unfractionated. Short GO terms include the annotations ‘membrane’, ‘integral to membrane’, ‘intrinsic to membrane’, ‘cell part’, but have no information as to subcellular localization.

Mentions: To identify cell surface proteins modulated by BCG infection, we used ‘plasma membrane profiling’ to compare infected with control cells. We fractionated peptides using high pH reversed phase liquid chromatography to increase the number of proteins quantified. We quantified 873 proteins, of which 559 had a Gene Ontology annotation ‘plasma membrane’ (PM), ‘cell surface’ (CS), ‘extracellular’ (XC), or ‘short GO’ (ShG, see methods) (70% of all quantified proteins with a GO annotation; Experiment A). To ensure all light-labeled skin contaminants were fully excluded, a ‘label swap’ unfractionated sample was also analyzed which identified 110 proteins, of which 93% (101 proteins) were annotated PM/CS/XC/ShG (Experiment B, Figure 1). We considered the overlap between Experiments A and B ‘high confidence’ data, which is displayed in Figures 2A,B; Supplementary Table S1. Full data is shown in Supplementary Table S2.


Plasma Membrane Profiling Reveals Upregulation of ABCA1 by Infected Macrophages Leading to Restriction of Mycobacterial Growth.

Long J, Basu Roy R, Zhang YJ, Antrobus R, Du Y, Smith DL, Weekes MP, Javid B - Front Microbiol (2016)

Gene ontology annotations of proteins identified by two or more peptides in each experiment. In Experiment A, peptides were fractionated by HpRp, and in Experiment B (label swap) unfractionated. Short GO terms include the annotations ‘membrane’, ‘integral to membrane’, ‘intrinsic to membrane’, ‘cell part’, but have no information as to subcellular localization.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940386&req=5

Figure 1: Gene ontology annotations of proteins identified by two or more peptides in each experiment. In Experiment A, peptides were fractionated by HpRp, and in Experiment B (label swap) unfractionated. Short GO terms include the annotations ‘membrane’, ‘integral to membrane’, ‘intrinsic to membrane’, ‘cell part’, but have no information as to subcellular localization.
Mentions: To identify cell surface proteins modulated by BCG infection, we used ‘plasma membrane profiling’ to compare infected with control cells. We fractionated peptides using high pH reversed phase liquid chromatography to increase the number of proteins quantified. We quantified 873 proteins, of which 559 had a Gene Ontology annotation ‘plasma membrane’ (PM), ‘cell surface’ (CS), ‘extracellular’ (XC), or ‘short GO’ (ShG, see methods) (70% of all quantified proteins with a GO annotation; Experiment A). To ensure all light-labeled skin contaminants were fully excluded, a ‘label swap’ unfractionated sample was also analyzed which identified 110 proteins, of which 93% (101 proteins) were annotated PM/CS/XC/ShG (Experiment B, Figure 1). We considered the overlap between Experiments A and B ‘high confidence’ data, which is displayed in Figures 2A,B; Supplementary Table S1. Full data is shown in Supplementary Table S2.

Bottom Line: The plasma membrane represents a critical interface between the internal and extracellular environments, and harbors multiple proteins key receptors and transporters that play important roles in restriction of intracellular infection.We quantified 559 PM proteins in BCG-infected THP-1 cells.One significantly upregulated cell-surface protein was the cholesterol transporter ABCA1.

View Article: PubMed Central - PubMed

Affiliation: Collaboration Innovation Centre for the Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University Beijing, China.

ABSTRACT
The plasma membrane represents a critical interface between the internal and extracellular environments, and harbors multiple proteins key receptors and transporters that play important roles in restriction of intracellular infection. We applied plasma membrane profiling, a technique that combines quantitative mass spectrometry with selective cell surface aminooxy-biotinylation, to Bacille Calmette-Guérin (BCG)-infected THP-1 macrophages. We quantified 559 PM proteins in BCG-infected THP-1 cells. One significantly upregulated cell-surface protein was the cholesterol transporter ABCA1. We showed that ABCA1 was upregulated on the macrophage cell-surface following infection with pathogenic mycobacteria and knockdown of ABCA1 resulted in increased mycobacterial survival within macrophages, suggesting that it may be a novel mycobacterial host-restriction factor.

No MeSH data available.


Related in: MedlinePlus