ProGP51

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ProGP ID ProGP51
Validation Status Characterized
Organism Information
Organism NameMycobacterium tuberculosis H37Rv
Domain Bacteria
Classification Family: Mycobacteriaceae
Suborder: Corynebacterineae
Order: Actinomycetales
Subclass: Actinobacteridae
Class: Actinobacteria
Division or phylum: "Actinobacteria"
Taxonomic ID (NCBI) 1773
Genome Sequence(s)
GenBank BX842578.1
EMBL BX842578
Organism Additional Information It is the causative agent of human tuberculosis. The pathogenesis is influenced by its lipoglycans and glycolipids (having a wide range of immunomodulatory activities), and a variety of its virulence factors and antigens.
Gene Information
Gene NameApa (Rv1860) or modD
NCBI Gene ID 885896
GenBank Gene Sequence 885896
Protein Information
Protein NameAlanine and proline-rich secreted protein Apa (50/55-kDa or 45 kDa MPT 32)
UniProtKB/SwissProt ID P9WIR7
NCBI RefSeq YP_177849.1
EMBL-CDSCAE55438.1
UniProtKB Sequence >sp|P9WIR6|APA_MYCTO Alanine and proline-rich secreted protein Apa OS=Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh) GN=apa PE=3 SV=1 MHQVDPNLTRRKGRLAALAIAAMASASLVTVAVPATANADPEPAPPVPTTAASPPSTAAA PPAPATPVAPPPPAAANTPNAQPGDPNAAPPPADPNAPPPPVIAPNAPQPVRIDNPVGGF SFALPAGWVESDAAHLDYGSALLSKTTGDPPFPGQPPPVANDTRIVLGRLDQKLYASAEA TDSKAAARLGSDMGEFYMPYPGTRINQETVSLDANGVSGSASYYEVKFSDPSKPNGQIWT GVIGSPAANAPDAGPPQRWFVVWLGTANNPVDKGAAKALAESIRPLVAPPPAPAPAPAEP APAPAPAGEVAPTPTTPTPQRTLPA
Sequence length 325 AA
Subcellular LocationSecreted
Function Immunodominant antigen. Elicits potent DTH (delayed-type hypersensitivity) responses in living-BCG-immunized guinea pigs..Stimulation of peripheral blood mononuclear cells from PPD positive individuals with Apa induces a Th1 type lymphoproliferative response with expansion of both CD4+ and CD8+ cells and increased IFN-γ production. 2) In Mtb, mannosylated nApa is a potential adhesin and has a role in host cell attachment, entry, and immune evasion, it induces a strong delayed-type hypersensitivity (DTH) in immunized animals, Elicits a mostly Th1 type of T-cell response in healthy humans; induces IFN-gamma production from CD4+ and CD8+ cells, Functions as an adhesin, binds to mouse macrophages via mannose residues
Glycosylation Status
Glycosylation Type O- (Thr) linked
Experimentally Validated Glycosite(s) in Full Length Protein(Signal peptide: 1-39) T49, T57, T66, T316, T313,T315
Experimentally Validated Glycosite(s ) in Mature ProteinT10, T18, T27, T277
Glycosite(s) Annotated Protein Sequence >sp|Q50906|APA_MYCTU Alanine and proline-rich secreted protein apa OS=Mycobacterium tuberculosis GN=apa PE=1 SV=1 MHQVDPNLTRRKGRLAALAIAAMASASLVTVAVPATANADPEPAPPVPT*(49)TAASPPST*(57)AAA PPAPAT*(66)PVAPPPPAAANTPNAQPGDPNAAPPPADPNAPPPPVIAPNAPQPVRIDNPVGGF SFALPAGWVESDAAHFDYGSALLSKTTGDPPFPGQPPPVANDTRIVLGRLDQKLYASAEA TDSKAAARLGSDMGEFYMPYPGTRINQETVSLDANGVSGSASYYEVKFSDPSKPNGQIWT GVIGSPAANAPDAGPPQRWFVVWLGTANNPVDKGAAKALAESIRPLVAPPPAPAPAPAEP APAPAPAGEVAPTPTT*(316)PTPQRTLPA
Sequence Around Glycosites (21 AA) ADPEPAPPVPTTAASPPSTAA
VPTTAASPPSTAAAPPAPATP
STAAAPPAPATPVAPPPPAAA
PAGEVAPTPTTPTPQRTLPA
Glycosite Sequence Logo
Technique(s) used for Glycosylation DetectionSchiff's staining, peroxidase-conjugated concanavalin A (ConA)-binding
Technique(s) used for Glycosylated Residue(s) Detection N-terminal amino acid sequencing coupled with fast atom bombardment-mass spectrometry (FAB-MS), Edman degradation on a gas-phase sequencer , FAB-MS
Protein Glycosylation- Implication The presence of the mannose residues on the Apa protein was essential for the antigenicity of the molecules in T-cell-dependent immune responses in vitro (proliferation assay) and in vivo (delayed type hypersensitivity or DTH reaction). The deglycosylated antigen was 10-fold less active than native molecules in eliciting DTH. Mannosylation of antigen leads to selective targeting and subsequent greater presentation by dendritic cells. 2) In Mtb,mannosylated nApa is a potential adhesin and has a role in host cell attachment, entry, and immune evasion
Glycan Information
Glycan Annotation Linkage: αMan-Thr.
34.6% sugar detected.
α-D-Manp(1→2)α-D-Manp (mannobiose), α-D-Manp (single mannose), α-D-Manp(1→2)α-D-Manp(1→2)α-D-Manp (mannotriose) are present. T10 and T18 are glycosylated with mannobiose, T27 with single mannose, and T277 with either of the three. The majority of the antigen species bear six, seven, or eight mannose residues (22, 24, and 17%, respectively), while others three, four, or five mannoses (5, 9, and 14%, respectively). Alpha-D-Man, mannobiose, or mannotriose, Thr residues at positions 10 and 18 were substituted a-D-Manp(132)a-D-Manp, position 27 was substituted with a single a-D-Manp, Thr-277 was substituted with either a-D-Manp, a-D-Manp(132)a-D-Manp, or a-D-Manp(132)a-D-Manp (132)a-D-Manp.
Technique(s) used for Glycan Identification GC-MS of partially methylated alditol acetates obtained by trifluoro acetic acid (TFA) hydrolysis of permethylated oligoglycosyl alditols which are released by β-elimination of the glycopeptides.
Protein Glycosylation linked (PGL) gene(s)
OST Gene NameProtein O mannosyltransferase
OST ProGT IDProGT10
Characterized Accessory Gene(s)Polyprenol-phosphate-mannose (PPM) synthase, Ppm1, is present. A second type of Ppm synthase (Rv3779 gene product) exclusive to slow-growing mycobacteria, is a membrane glycosyltransferase. It mannosylates polyprenyl-phosphates directly from GDP-mannose.
Accessory Gene(s)Progt IDProGT10.1
Additional CommentSec-mediated translocation influences the O-mannosylation. Ppm1 does not discriminate between polyprenol substrates with variable chain lengths and saturation of the isoprene units.
Glycosylation sites have been found to be located within proline-rich domains (or Thr-rich sequences) near the N-terminus and the C-terminus.
In a cell-free assay, the M. smegmatis mannosyltransferase activity in membrane and cell wall fraction has been shown to catalyze transfer of radiolabeled mannose from GDP-[14C]mannose to peptide acceptors. The acceptors consisted of Thr-rich sequences from M. tuberculosis 45 kDa antigen (Ref. no. 3).
The 45/47 kDa antigen (Apa) has also been glycosylated in Streptomyces lividans. The 45- and 47-kDa represent two glycoforms of the antigen (Ref. no. 2).
Literature
Year of Identification1989
Year of Identification Month Wise1989.09.01
Year of Validation 1996
ReferenceScherman, H., Kaur, D., Pham, H., Skovierova, H., Jackson, M. and Brennan, P.J. (2009) Identification of a polyprenylphosphomannosyl synthase involved in the synthesis of mycobacterial mannosides. J Bacteriol, 191, 6769-6772. [PubMed: 19717608]
AuthorScherman, H., Kaur, D., Pham, H., Skovierova, H., Jackson, M. and Brennan, P.J.
Research GroupDepartment of Microbiology, Colorado State University, Fort Collins, Colorado 80523-1682, USA.
Corresponding Author Brennan, P.J
ContactDepartment of Microbiology, Colorado State University, Fort Collins, Colorado 80523-1682, USA.
ReferenceLara, M., Servin-Gonzalez, L., Singh, M., Moreno, C., Cohen, I., Nimtz, M. and Espitia, C. (2004) Expression, secretion, and glycosylation of the 45- and 47-kDa glycoprotein of Mycobacterium tuberculosis in Streptomyces lividans. Appl Environ Microbiol, 70, 679-685. [PubMed: 14766542]
Author Lara, M., Servin-Gonzalez, L., Singh, M., Moreno, C., Cohen, I., Nimtz, M. Espitia, C.
Research GroupDepartment of Immunology, Institute of Biomedical Research, National Autonomous University of Mexico, Mexico D.F., Mexico.
Corresponding Author Espitia, C.
ContactDepartment of Immunology, Institute of Biomedical Research, National Autonomous University of Mexico, Mexico D.F., Mexico.
ReferenceLara, M., Servin-Gonzalez, L., Singh, M., Moreno, C., Cohen, I., Nimtz, M. and Espitia, C. (2004) Expression, secretion, and glycosylation of the 45- and 47-kDa glycoprotein of Mycobacterium tuberculosis in Streptomyces lividans. Appl Environ Microbiol, 70, 679-685. [PubMed: 14766542]
Author Lara, M., Servin-Gonzalez, L., Singh, M., Moreno, C., Cohen, I., Nimtz, M. Espitia, C.
Research GroupDepartment of Immunology, Institute of Biomedical Research, National Autonomous University of Mexico, Mexico D.F., Mexico
Corresponding Author Espitia, C.
ContactDepartment of Immunology, Institute of Biomedical Research, National Autonomous University of Mexico, Mexico D.F., Mexico
Reference Cooper, H.N., Gurcha, S.S., Nigou, J., Brennan, P.J., Belisle, J.T., Besra, G.S. and Young, D. (2002) Characterization of mycobacterial protein glycosyltransferase activity using synthetic peptide acceptors in a cell-free assay. Glycobiology, 12, 427-434. [PubMed: 12122024]
Author Cooper, H.N., Gurcha, S.S., Nigou, J., Brennan, P.J., Belisle, J.T., Besra, G.S. Young, D.
Research GroupCentre for Molecular Microbiology and Infection, Imperial College of Science, Technology and Medicine, South Kensington, London, SW7 2AZ, England.
Corresponding Author Young, D.
ContactCentre for Molecular Microbiology and Infection, Imperial College of Science, Technology and Medicine, South Kensington, London, SW7 2AZ, England.
Reference Horn, C., Namane, A., Pescher, P., Riviere, M., Romain, F., Puzo, G., Barzu, O. and Marchal, G. (1999) Decreased capacity of recombinant 45/47-kDa molecules (Apa) of Mycobacterium tuberculosis to stimulate T lymphocyte responses related to changes in their mannosylation pattern. J Biol Chem, 274, 32023-32030. [PubMed: 10542234]
Author Horn, C., Namane, A., Pescher, P., Riviere, M., Romain, F., Puzo, G., Barzu, O. Marchal, G.
Research GroupDepartment of Pathophysiology of Infection, Pasteur Institute, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.
Corresponding Author Marchal, G.
ContactDepartment of Pathophysiology of Infection, Pasteur Institute, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.
Reference Romain, F., Horn, C., Pescher, P., Namane, A., Riviere, M., Puzo, G., Barzu, O. and Marchal, G. (1999) Deglycosylation of the 45/47-kilodalton antigen complex of Mycobacterium tuberculosis decreases its capacity to elicit in vivo or in vitro cellular immune responses. Infect Immun, 67, 5567-5572. [PubMed: 10531201]
Author Romain, F., Horn, C., Pescher, P., Namane, A., Riviere, M., Puzo, G., Barzu, O. Marchal, G.
Research GroupUnit of Physiopathology of Infection, Institut Pasteur, 75724 Paris Cedex 15, France.
Corresponding Author Marchal, G.
ContactUnit of Physiopathology of Infection, Institut Pasteur, 75724 Paris Cedex 15, France.
Reference Dobos, K.M., Khoo, K.H., Swiderek, K.M., Brennan, P.J. and Belisle, J.T. (1996) Definition of the full extent of glycosylation of the 45-kilodalton glycoprotein of Mycobacterium tuberculosis. J Bacteriol, 178, 2498-2506. [PubMed: 8626314]
Author Dobos, K.M., Khoo, K.H., Swiderek, K.M., Brennan, P.J. and Belisle, J.T.
Research GroupDepartment of Microbiology, Colorado State University, Fort Collins 80523, USA.
Corresponding Author Belisle, J.T.
ContactDepartment of Microbiology, Colorado State University, Fort Collins 80523, USA.
Reference Espitia, C., Espinosa, R., Saavedra, R., Mancilla, R., Romain, F., Laqueyrerie, A. and Moreno, C. (1995) Antigenic and structural similarities between Mycobacterium tuberculosis 50- to 55-kilodalton and Mycobacterium bovis BCG 45- to 47-kilodalton antigens. Infect Immun, 63, 580-584. [PubMed: 7822025]
Author Espitia, C., Espinosa, R., Saavedra, R., Mancilla, R., Romain, F., Laqueyrerie, A. and Moreno, C.
Research GroupTuberculosis and Related Infections Unit, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.
Corresponding Author Moreno, C.
ContactTuberculosis and Related Infections Unit, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.
Reference Espitia, C. and Mancilla, R. (1989) Identification, isolation and partial characterization of Mycobacterium tuberculosis glycoprotein antigens. Clin Exp Immunol, 77, 378-383. [PubMed: 2478323]
Author Espitia, C. and Mancilla, R.
Research GroupDepartment of Immunology, National Autonomous University of Mexico, D.F.
Corresponding Author Mancilla, R.
ContactDepartment of Immunology, National Autonomous University of Mexico, D.F.
ReferenceNandakumar S1, Kannanganat S, Dobos KM, Lucas M, Spencer JS, Fang S, McDonald MA, Pohl J, Birkness K, Chamcha V, Ramirez MV, Plikaytis BB, Posey JE, Amara RR, Sable SB.(2013) O-mannosylation of the Mycobacterium tuberculosis adhesin Apa is crucial for T cell antigenicity during infection but is expendable for protection. PLoS Pathog. 2013;9(10):e1003705. doi: 10.1371/journal.ppat.1003705. Epub 2013 Oct 10.
AuthorNandakumar S1, Kannanganat S, Dobos KM, Lucas M, Spencer JS, Fang S, McDonald MA, Pohl J, Birkness K, Chamcha V, Ramirez MV, Plikaytis BB, Posey JE, Amara RR, Sable SB.
Research GroupDivision of Tuberculosis Elimination, National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
Corresponding Author Sable SB
ContactDivision of Tuberculosis Elimination, National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.