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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

Flow cytometry histograms of a selection of proteins with altered cell surface expression following BCG infection of THP-1 macrophages. Macrophages were infected with BCG-GFP (MOI: 5:1) for 4 h and then analyzed 48 h following infection by flow cytometry. Infected macrophages (GFPHI) and uninfected macrophages (GFPLO) were compared by mean fluorescent intensity (MFI).
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Figure 3: Flow cytometry histograms of a selection of proteins with altered cell surface expression following BCG infection of THP-1 macrophages. Macrophages were infected with BCG-GFP (MOI: 5:1) for 4 h and then analyzed 48 h following infection by flow cytometry. Infected macrophages (GFPHI) and uninfected macrophages (GFPLO) were compared by mean fluorescent intensity (MFI).

Mentions: We selected a subset of PM proteins whose expression was significantly affected by BCG infection and for which commercially available antibodies were available for flow cytometry (Figure 3). We confirmed upregulation of the most significantly upregulated high confidence proteins CD14 and ATP-binding cassette sub-family A member 1 (ABCA1) (Figures 2 and 3), in addition to SLAM Family Member 7 (SLAMF7) (Figure 3; 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)

Flow cytometry histograms of a selection of proteins with altered cell surface expression following BCG infection of THP-1 macrophages. Macrophages were infected with BCG-GFP (MOI: 5:1) for 4 h and then analyzed 48 h following infection by flow cytometry. Infected macrophages (GFPHI) and uninfected macrophages (GFPLO) were compared by mean fluorescent intensity (MFI).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4940386&req=5

Figure 3: Flow cytometry histograms of a selection of proteins with altered cell surface expression following BCG infection of THP-1 macrophages. Macrophages were infected with BCG-GFP (MOI: 5:1) for 4 h and then analyzed 48 h following infection by flow cytometry. Infected macrophages (GFPHI) and uninfected macrophages (GFPLO) were compared by mean fluorescent intensity (MFI).
Mentions: We selected a subset of PM proteins whose expression was significantly affected by BCG infection and for which commercially available antibodies were available for flow cytometry (Figure 3). We confirmed upregulation of the most significantly upregulated high confidence proteins CD14 and ATP-binding cassette sub-family A member 1 (ABCA1) (Figures 2 and 3), in addition to SLAM Family Member 7 (SLAMF7) (Figure 3; 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