Acylation state of the phosphatidylinositol hexamannosides from Mycobacterium bovis bacillus Calmette Guerin and mycobacterium tuberculosis H37Rv and its implication in Toll-like receptor response.
The dimannoside (PIM2) and hexamannoside (PIM6) phosphatidyl-myo-inositol mannosides are the two most abundant classes of PIM found in Mycobacterium bovis bacillus Calmette Guérin, Mycobacterium tuberculosis H37Rv, and Mycobacterium smegmatis 607. Recently, these long known molecules received a renewed interest due to the fact that PIM2 constitute the anchor motif of an important constituent of the mycobacterial cell wall, the lipoarabinomannans (LAM), and that both LAM (phosphoinositol-capped LAM) and PIM are agonists of Toll-like receptor 2 (TLR2), a pattern recognition receptor involved in innate immunity. Due to the biological importance of these molecules, the chemical structure of PIM was revisited. The structure of PIM2 was recently published (Gilleron, M., Ronet, C., Mempel, M., Monsarrat, B., Gachelin, G., and Puzo, G. (2001) J. Biol. Chem. 276, 34896-34904). Here we report the purification and molecular characterization of PIM6 in their native form. For the first time, four acyl forms of this molecule have been purified, using hydrophobic interaction chromatography. Mono- to tetra-acylated molecules were identified in M. bovis bacillus Calmette Guérin, M. tuberculosis H37Rv, and M. smegmatis 607 using a sophisticated combination of analytical tools, including matrix-assisted laser desorption/ionization-time of flight-mass spectrometry and two-dimensional homo- and heteronuclear NMR spectroscopy. These experiments revealed that the major acyl forms are similar to the ones described for PIM2. Finally, we show that PIM6, like PIM2, activate primary macrophages to secrete TNF-alpha through TLR2, irrespective of their acylation pattern, and that they signal through the adaptor MyD88.
Gilleron
Martine
M
Institut de Pharmacologie et de Biologie Structurale du CNRS, 205 Route de Narbonne, 31077 Toulouse Cedex, France. Martine.Gilleron@ipbs.fr
Quesniaux
Valérie F J
VF
Puzo
Germain
G
D020816:Amino Acid Motifs; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D002855:Chromatography, Thin Layer; D005227:Fatty Acids / Q000737:chemistry; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D009682:Magnetic Resonance Spectroscopy; D008351:Mannans / Q000737:chemistry*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008956:Models, Chemical; D009163:Mycobacterium bovis / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D051195:Toll-Like Receptor 2; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D005227:Fatty Acids; D008351:Mannans; D008562:Membrane Glycoproteins; D010716:Phosphatidylinositols; D011956:Receptors, Cell Surface; D051195:Toll-Like Receptor 2; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; C054977:phosphatidylinositol dimannoside