UniProt functional annotation for P9WK45



	UniProt functional annotation for P9WK45

UniProt code: P9WK45.

Organism: Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).

Taxonomy: Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae; Mycobacterium; Mycobacterium tuberculosis complex.

Function: Probably helps membrane protein Rv1410c (P55) transport triacylglycerides (TAG) across the inner cell membrane into the periplasm; TAG probably regulates lipid metabolism and growth regulation (PubMed:26751071). Binds TAG and transfers it between lipid bilayers, probably to the outer membrane in vivo (PubMed:26751071). Binds di- and triacylated phosphatidyl-myo-inositol mannosides (PIMs), and glycolipid lipoglycan modulins lipoarabinomannan (LAM) and lipomannan (LM), facilitating their recognition by TLR2 (PubMed:20694006, PubMed:25356793). Binds LM > PIM6 > ManLAM > PI-LAM > PIM2 (mannose-capped LAM and phospho-myo-inositol-capped LAM, E.coli expressed without acyl-groups); deacylated LM and LAM also bind to this protein via their mannose moieties, showing LprG has at least 2 different ways to bind glycolipids (PubMed:25356793). Binds triacylglycerides (TAG) in the same cavity, is able to transfer TAG between lipid bilayers (PubMed:26751071). Overexpression of LprG and Rv1410c leads to increased levels of TAG in the culture medium (PubMed:26751071). Required for Rv1410c-mediated export of drugs (PubMed:18156250, PubMed:21762531). Required, probably with Rv1410c, for normal surface localization of LAM (PubMed:25232742). {ECO:0000269|PubMed:18156250, ECO:0000269|PubMed:20694006, ECO:0000269|PubMed:21762531, ECO:0000269|PubMed:25232742, ECO:0000269|PubMed:25356793, ECO:0000269|PubMed:26751071}.

Function: A host TLR2 agonist (toll-like receptor), shown experimentally for human and mouse (PubMed:19362712). Inhibits primary human macrophage MHC-II Ag processing via TLR2 (PubMed:15294983). Both lipidated and nonlipidated protein act as TLR2 agonists in antigen- presenting cells, although lipidated protein is more efficient (PubMed:20694006). In resting human CD4+ T-cells lipidated but not nonlipidated protein is a costimulatory ligand (with anti-CD3 and anti- CD28) for T-cell proliferation and IFN-gamma and IL-2 production, leading to increased expression of early T-cell activation markers, TLR2 and NFKB3 phosphorylation (PubMed:21078852). Human CD4+ T-cells use TLR1/TLR2 heterodimers to respond to this and probably other mycobacterial lipoproteins (PubMed:21078852). Able to stimulate proliferation of CD4+ T-cells derived from a human leprosy patient following protein processing/presentation by MHC class II molecules in peripheral blood mononuclear cells (PubMed:18424702). Requires both host TLR1 and TLR2 as coreceptors to elicit host response in mouse, although TLR6 may play a redundant role, has a partial requirement for CD14 as an accessory receptor (PubMed:19362712). {ECO:0000269|PubMed:15294983, ECO:0000269|PubMed:18424702, ECO:0000269|PubMed:19362712, ECO:0000269|PubMed:20694006, ECO:0000269|PubMed:21078852}.

Subcellular location: Cell inner membrane {ECO:0000255|PROSITE- ProRule:PRU00303, ECO:0000269|PubMed:21762531, ECO:0000305}; Lipid- anchor {ECO:0000255|PROSITE-ProRule:PRU00303}. Secreted, cell wall {ECO:0000269|PubMed:21762531}. Secreted {ECO:0000269|PubMed:21762531}. Cell surface {ECO:0000269|PubMed:25041568}. Note=Present in extracytoplasmic vesicles (PubMed:21364279). Immunoelectron microscopy indicates this protein is close to the cell surface (PubMed:25041568). {ECO:0000269|PubMed:21364279, ECO:0000269|PubMed:25041568}.

Domain: Forms a U-shaped beta-half-barrel with a small hydrophobic cavity (1500 Angstroms (3)) which holds a triacylated PIM in 1 crystal structure; the 3 acyl chains are within the cavity while the sugar moieties bind to the protein surface (PubMed:20694006). In the structure bound to triacylglycerides (TAG) 2 of the 3 acyl chains are buried in the cavity, the third is solvent exposed (PubMed:26751071). A flexible lid region may move to accommodate different TAG molecules (PubMed:26751071). Fragments of the mature protein (residues 81-100, 141-160 and 218-236) prevent uptake of M.tuberculosis by a human macrophage-like cell line; lesser effects are seen on bacterial uptake by a human lung epithelial cell line (PubMed:25041568). {ECO:0000269|PubMed:20694006, ECO:0000269|PubMed:25041568, ECO:0000269|PubMed:26751071}.

Ptm: Modified by Lgt on Cys-27 with an S-linked diacylglyceral, signal peptide is removed by LspA, Cys-27 is further modifed with a fatty acid on its amino group by Lnt yielding a triacylated protein (Probable). Probably glycosylated, which is required for T-cell activation (PubMed:18424702). {ECO:0000250|UniProtKB:P9WK47, ECO:0000269|PubMed:18424702, ECO:0000305|PubMed:20694006}.

Disruption phenotype: A single deletion mutant leads to loss of expression of efflux pump Rv1410c due to polar effects; in infected BALB/c mice 1.5 and 2.5 log decrease in bacterial load 15 and 35 days after infection (PubMed:14998516). The single mutant increases sensitivity to malachite green, sodium dodecyl sulfate (SDS), isoniazid, ethambutal and ethidium bromide, alters the permeability of the cell wall; both genes of the operon are required to fully restore the phenotypes (PubMed:21762531). Single deletion mutant (probably without Rv1410c) has decreased surface-exposed glycolipid lipoarabinomannan (LAM), although cellular LAM, LM and PIM content is normal (PubMed:25232742, PubMed:25356793). Disruption of either Rv1410c or the lrpG-Rv1410c operon leads to increased levels of many triacylglyceride (TAG) alkylforms; up to 100-fold increase depending on the exact TAG form (PubMed:26751071). It also forms smaller colonies on agar (PubMed:25232742). Loss of surface LAM has several consequences; bacteria enter mouse macrophages with reduced efficiency and block mouse macrophage phagosome-lysosome fusion less efficiently than wild- type (PubMed:25232742). Reduced efficiency of mouse macrophage phagosome-lysosome fusion was seen in another study (PubMed:25356793). C57BL/6 mice infected with mutant bacteria have 10-fold less bacterial burden after 10 days and about 2700-fold less burden after 70 days; attenuation of mutant is not rescued in macrophages impaired for reactive oxygen or nitrogen generation (disruption of Ncf1 or iNOS) (PubMed:25232742). {ECO:0000269|PubMed:14998516, ECO:0000269|PubMed:21762531, ECO:0000269|PubMed:25232742, ECO:0000269|PubMed:25356793, ECO:0000269|PubMed:26751071}.

Miscellaneous: Bacterial LAM blocks host cell phagosome-lysosome fusion and is one way in which M.tuberculosis evades the host immune system. {ECO:0000305|PubMed:25232742, ECO:0000305|PubMed:25356793}.

Miscellaneous: Triacylglycerides accumulate in lipid droplets in the cytoplasm of M.tuberculosis stationary phase and dormant bacteria, and are used as an energy source during starvation (PubMed:26751071). {ECO:0000305|PubMed:26751071}.

Similarity: Belongs to the LppX/LprAFG lipoprotein family. {ECO:0000305}.

Annotations taken from UniProtKB at the EBI.


Organism:		Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
Taxonomy:		Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae; Mycobacterium; Mycobacterium tuberculosis complex.



Function:		Probably helps membrane protein Rv1410c (P55) transport triacylglycerides (TAG) across the inner cell membrane into the periplasm; TAG probably regulates lipid metabolism and growth regulation (PubMed:26751071). Binds TAG and transfers it between lipid bilayers, probably to the outer membrane in vivo (PubMed:26751071). Binds di- and triacylated phosphatidyl-myo-inositol mannosides (PIMs), and glycolipid lipoglycan modulins lipoarabinomannan (LAM) and lipomannan (LM), facilitating their recognition by TLR2 (PubMed:20694006, PubMed:25356793). Binds LM > PIM6 > ManLAM > PI-LAM > PIM2 (mannose-capped LAM and phospho-myo-inositol-capped LAM, E.coli expressed without acyl-groups); deacylated LM and LAM also bind to this protein via their mannose moieties, showing LprG has at least 2 different ways to bind glycolipids (PubMed:25356793). Binds triacylglycerides (TAG) in the same cavity, is able to transfer TAG between lipid bilayers (PubMed:26751071). Overexpression of LprG and Rv1410c leads to increased levels of TAG in the culture medium (PubMed:26751071). Required for Rv1410c-mediated export of drugs (PubMed:18156250, PubMed:21762531). Required, probably with Rv1410c, for normal surface localization of LAM (PubMed:25232742). {ECO:0000269\|PubMed:18156250, ECO:0000269\|PubMed:20694006, ECO:0000269\|PubMed:21762531, ECO:0000269\|PubMed:25232742, ECO:0000269\|PubMed:25356793, ECO:0000269\|PubMed:26751071}.



Function:		A host TLR2 agonist (toll-like receptor), shown experimentally for human and mouse (PubMed:19362712). Inhibits primary human macrophage MHC-II Ag processing via TLR2 (PubMed:15294983). Both lipidated and nonlipidated protein act as TLR2 agonists in antigen- presenting cells, although lipidated protein is more efficient (PubMed:20694006). In resting human CD4+ T-cells lipidated but not nonlipidated protein is a costimulatory ligand (with anti-CD3 and anti- CD28) for T-cell proliferation and IFN-gamma and IL-2 production, leading to increased expression of early T-cell activation markers, TLR2 and NFKB3 phosphorylation (PubMed:21078852). Human CD4+ T-cells use TLR1/TLR2 heterodimers to respond to this and probably other mycobacterial lipoproteins (PubMed:21078852). Able to stimulate proliferation of CD4+ T-cells derived from a human leprosy patient following protein processing/presentation by MHC class II molecules in peripheral blood mononuclear cells (PubMed:18424702). Requires both host TLR1 and TLR2 as coreceptors to elicit host response in mouse, although TLR6 may play a redundant role, has a partial requirement for CD14 as an accessory receptor (PubMed:19362712). {ECO:0000269\|PubMed:15294983, ECO:0000269\|PubMed:18424702, ECO:0000269\|PubMed:19362712, ECO:0000269\|PubMed:20694006, ECO:0000269\|PubMed:21078852}.


Subcellular location:		Cell inner membrane {ECO:0000255\|PROSITE- ProRule:PRU00303, ECO:0000269\|PubMed:21762531, ECO:0000305}; Lipid- anchor {ECO:0000255\|PROSITE-ProRule:PRU00303}. Secreted, cell wall {ECO:0000269\|PubMed:21762531}. Secreted {ECO:0000269\|PubMed:21762531}. Cell surface {ECO:0000269\|PubMed:25041568}. Note=Present in extracytoplasmic vesicles (PubMed:21364279). Immunoelectron microscopy indicates this protein is close to the cell surface (PubMed:25041568). {ECO:0000269\|PubMed:21364279, ECO:0000269\|PubMed:25041568}.
Domain:		Forms a U-shaped beta-half-barrel with a small hydrophobic cavity (1500 Angstroms (3)) which holds a triacylated PIM in 1 crystal structure; the 3 acyl chains are within the cavity while the sugar moieties bind to the protein surface (PubMed:20694006). In the structure bound to triacylglycerides (TAG) 2 of the 3 acyl chains are buried in the cavity, the third is solvent exposed (PubMed:26751071). A flexible lid region may move to accommodate different TAG molecules (PubMed:26751071). Fragments of the mature protein (residues 81-100, 141-160 and 218-236) prevent uptake of M.tuberculosis by a human macrophage-like cell line; lesser effects are seen on bacterial uptake by a human lung epithelial cell line (PubMed:25041568). {ECO:0000269\|PubMed:20694006, ECO:0000269\|PubMed:25041568, ECO:0000269\|PubMed:26751071}.
Ptm:		Modified by Lgt on Cys-27 with an S-linked diacylglyceral, signal peptide is removed by LspA, Cys-27 is further modifed with a fatty acid on its amino group by Lnt yielding a triacylated protein (Probable). Probably glycosylated, which is required for T-cell activation (PubMed:18424702). {ECO:0000250\|UniProtKB:P9WK47, ECO:0000269\|PubMed:18424702, ECO:0000305\|PubMed:20694006}.
Disruption phenotype:		A single deletion mutant leads to loss of expression of efflux pump Rv1410c due to polar effects; in infected BALB/c mice 1.5 and 2.5 log decrease in bacterial load 15 and 35 days after infection (PubMed:14998516). The single mutant increases sensitivity to malachite green, sodium dodecyl sulfate (SDS), isoniazid, ethambutal and ethidium bromide, alters the permeability of the cell wall; both genes of the operon are required to fully restore the phenotypes (PubMed:21762531). Single deletion mutant (probably without Rv1410c) has decreased surface-exposed glycolipid lipoarabinomannan (LAM), although cellular LAM, LM and PIM content is normal (PubMed:25232742, PubMed:25356793). Disruption of either Rv1410c or the lrpG-Rv1410c operon leads to increased levels of many triacylglyceride (TAG) alkylforms; up to 100-fold increase depending on the exact TAG form (PubMed:26751071). It also forms smaller colonies on agar (PubMed:25232742). Loss of surface LAM has several consequences; bacteria enter mouse macrophages with reduced efficiency and block mouse macrophage phagosome-lysosome fusion less efficiently than wild- type (PubMed:25232742). Reduced efficiency of mouse macrophage phagosome-lysosome fusion was seen in another study (PubMed:25356793). C57BL/6 mice infected with mutant bacteria have 10-fold less bacterial burden after 10 days and about 2700-fold less burden after 70 days; attenuation of mutant is not rescued in macrophages impaired for reactive oxygen or nitrogen generation (disruption of Ncf1 or iNOS) (PubMed:25232742). {ECO:0000269\|PubMed:14998516, ECO:0000269\|PubMed:21762531, ECO:0000269\|PubMed:25232742, ECO:0000269\|PubMed:25356793, ECO:0000269\|PubMed:26751071}.
Miscellaneous:		Bacterial LAM blocks host cell phagosome-lysosome fusion and is one way in which M.tuberculosis evades the host immune system. {ECO:0000305\|PubMed:25232742, ECO:0000305\|PubMed:25356793}.
Miscellaneous:		Triacylglycerides accumulate in lipid droplets in the cytoplasm of M.tuberculosis stationary phase and dormant bacteria, and are used as an energy source during starvation (PubMed:26751071). {ECO:0000305\|PubMed:26751071}.
Similarity:		Belongs to the LppX/LprAFG lipoprotein family. {ECO:0000305}.