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Structural characterization and functional properties of a novel lipomannan variant isolated from a Corynebacterium glutamicum pimB' mutant.

Mishra AK, Klein C, Gurcha SS, Alderwick LJ, Babu P, Hitchen PG, Morris HR, Dell A, Besra GS, Eggeling L - Antonie Van Leeuwenhoek (2008)

Bottom Line: The disruption of NCgl2106 in C. glutamicum resulted in a mutant devoid of monoacylated phosphatidyl-myo-inositol dimannoside (Ac(1)PIM(2)) resulting in the accumulation of Ac(1)PIM(1) and cessation of phosphatidyl-myo-inositol (PI) based lipomannan (Cg-LM, now also termed 'Cg-LM-A') and lipoarabinomannan (Cg-LAM) biosynthesis.The deletion of NCgl2106 in C. glutamicum has now provided a useful strain, in addition with a deletion mutant of NCgl0452 in C. glutamicum for the purification of Cg-LM-A and Cg-LM-B.Interestingly, both Cg-LM species induced a similar production of TNF-alpha by a human macrophage cell line suggesting that the phospho-myo-inositol residue of the PI-anchor does not play a key role in lipoglycan pro-inflammatory activity.

View Article: PubMed Central - PubMed

Affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.

ABSTRACT
The genus Corynebacterium is part of the phylogenetic group nocardioform actinomycetes, which also includes the genus Mycobacterium. Members of this phylogenetic group have a characteristic cell envelope structure, which is dominated by complex lipids and amongst these, lipoglycans are of particular interest. The disruption of NCgl2106 in C. glutamicum resulted in a mutant devoid of monoacylated phosphatidyl-myo-inositol dimannoside (Ac(1)PIM(2)) resulting in the accumulation of Ac(1)PIM(1) and cessation of phosphatidyl-myo-inositol (PI) based lipomannan (Cg-LM, now also termed 'Cg-LM-A') and lipoarabinomannan (Cg-LAM) biosynthesis. Interestingly, SDS-analysis of the lipoglycan fraction from the mutant revealed the synthesis of a single novel lipoglycan, now termed 'Cg-LM-B'. Further chemical analyses established the lipoglycan possessed an alpha-D: -glucopyranosyluronic acid-(1 --> 3)-glycerol (GlcAGroAc(2)) based anchor which was then further glycosylated by 8-22 mannose residues, with Man(12-20)GlcAGroAC(2) molecular species being the most abundant, to form a novel lipomannan structure (Cg-LM-B). The deletion of NCgl2106 in C. glutamicum has now provided a useful strain, in addition with a deletion mutant of NCgl0452 in C. glutamicum for the purification of Cg-LM-A and Cg-LM-B. Interestingly, both Cg-LM species induced a similar production of TNF-alpha by a human macrophage cell line suggesting that the phospho-myo-inositol residue of the PI-anchor does not play a key role in lipoglycan pro-inflammatory activity.

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TNF-α production by human macrophage cell line in response to Cg-LMs. Cg-LM-A was isolated and purified as described previously (Tatituri et al. 2007) and Cg-LM-B (this study) were tested at 5 (grey bars) and 15 (black bars) μg/ml
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Fig4: TNF-α production by human macrophage cell line in response to Cg-LMs. Cg-LM-A was isolated and purified as described previously (Tatituri et al. 2007) and Cg-LM-B (this study) were tested at 5 (grey bars) and 15 (black bars) μg/ml

Mentions: PI-anchored lipoglycans, and most particularly LMs, isolated from other actinomycete genera, including mycobacteria, have previously been shown to induce cytokine production by phagocytic cells (Garton et al. 2002; Gibson et al. 2004; Quesniaux et al. 2004). We thus investigated the potency of both types of Cg-LMs to stimulate the release of TNF-α using a human macrophage cell line. As expected, the PI-anchored LM, Cg-LM-A, elicited a dose-dependent production of the cytokine (Fig. 4). However interestingly, the GlcAGroAc2-anchored LM, Cg-LM-B, was also found to be as stimulatory as the PI-anchored LM, suggesting that the phospho-myo-inositol unit of the anchor in LM does not play a key role in lipoglycan pro-inflammatory activity.Fig. 4


Structural characterization and functional properties of a novel lipomannan variant isolated from a Corynebacterium glutamicum pimB' mutant.

Mishra AK, Klein C, Gurcha SS, Alderwick LJ, Babu P, Hitchen PG, Morris HR, Dell A, Besra GS, Eggeling L - Antonie Van Leeuwenhoek (2008)

TNF-α production by human macrophage cell line in response to Cg-LMs. Cg-LM-A was isolated and purified as described previously (Tatituri et al. 2007) and Cg-LM-B (this study) were tested at 5 (grey bars) and 15 (black bars) μg/ml
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2480597&req=5

Fig4: TNF-α production by human macrophage cell line in response to Cg-LMs. Cg-LM-A was isolated and purified as described previously (Tatituri et al. 2007) and Cg-LM-B (this study) were tested at 5 (grey bars) and 15 (black bars) μg/ml
Mentions: PI-anchored lipoglycans, and most particularly LMs, isolated from other actinomycete genera, including mycobacteria, have previously been shown to induce cytokine production by phagocytic cells (Garton et al. 2002; Gibson et al. 2004; Quesniaux et al. 2004). We thus investigated the potency of both types of Cg-LMs to stimulate the release of TNF-α using a human macrophage cell line. As expected, the PI-anchored LM, Cg-LM-A, elicited a dose-dependent production of the cytokine (Fig. 4). However interestingly, the GlcAGroAc2-anchored LM, Cg-LM-B, was also found to be as stimulatory as the PI-anchored LM, suggesting that the phospho-myo-inositol unit of the anchor in LM does not play a key role in lipoglycan pro-inflammatory activity.Fig. 4

Bottom Line: The disruption of NCgl2106 in C. glutamicum resulted in a mutant devoid of monoacylated phosphatidyl-myo-inositol dimannoside (Ac(1)PIM(2)) resulting in the accumulation of Ac(1)PIM(1) and cessation of phosphatidyl-myo-inositol (PI) based lipomannan (Cg-LM, now also termed 'Cg-LM-A') and lipoarabinomannan (Cg-LAM) biosynthesis.The deletion of NCgl2106 in C. glutamicum has now provided a useful strain, in addition with a deletion mutant of NCgl0452 in C. glutamicum for the purification of Cg-LM-A and Cg-LM-B.Interestingly, both Cg-LM species induced a similar production of TNF-alpha by a human macrophage cell line suggesting that the phospho-myo-inositol residue of the PI-anchor does not play a key role in lipoglycan pro-inflammatory activity.

View Article: PubMed Central - PubMed

Affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.

ABSTRACT
The genus Corynebacterium is part of the phylogenetic group nocardioform actinomycetes, which also includes the genus Mycobacterium. Members of this phylogenetic group have a characteristic cell envelope structure, which is dominated by complex lipids and amongst these, lipoglycans are of particular interest. The disruption of NCgl2106 in C. glutamicum resulted in a mutant devoid of monoacylated phosphatidyl-myo-inositol dimannoside (Ac(1)PIM(2)) resulting in the accumulation of Ac(1)PIM(1) and cessation of phosphatidyl-myo-inositol (PI) based lipomannan (Cg-LM, now also termed 'Cg-LM-A') and lipoarabinomannan (Cg-LAM) biosynthesis. Interestingly, SDS-analysis of the lipoglycan fraction from the mutant revealed the synthesis of a single novel lipoglycan, now termed 'Cg-LM-B'. Further chemical analyses established the lipoglycan possessed an alpha-D: -glucopyranosyluronic acid-(1 --> 3)-glycerol (GlcAGroAc(2)) based anchor which was then further glycosylated by 8-22 mannose residues, with Man(12-20)GlcAGroAC(2) molecular species being the most abundant, to form a novel lipomannan structure (Cg-LM-B). The deletion of NCgl2106 in C. glutamicum has now provided a useful strain, in addition with a deletion mutant of NCgl0452 in C. glutamicum for the purification of Cg-LM-A and Cg-LM-B. Interestingly, both Cg-LM species induced a similar production of TNF-alpha by a human macrophage cell line suggesting that the phospho-myo-inositol residue of the PI-anchor does not play a key role in lipoglycan pro-inflammatory activity.

Show MeSH
Related in: MedlinePlus