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ProGT ID ProGT10
Validation StatusCharacterized
ProGT Pathway
Organism Information
Organism NameMycobacterium tuberculosis H37Rv ATCC 25618
Clinical ImplicationPathogenic
DomainBacteria
PhylumActinobacteria
ClassificationFamily: Mycobacteriaceae
Suborder: Corynebacterineae
Order: Actinomycetales
Subclass: Actinobacteridae
Class: Actinobacteria
Division or phylum: "Actinobacteria"
Taxonomic ID (NCBI)83332
Genome Sequence(s)
Gene BankNC_000962.3
EMBLAL123456
Gene Information
NCBI Gene ID887882
Protein information
UniProtKB/ SwissProt IDP9WN05
NCBI Ref SeqNP_215518.1
UniProtKB Sequence>sp|P9WN05|PMT_MYCTU Probable dolichyl-phosphate-mannose--protein mannosyltransferase OS=Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv) GN=pmt PE=1 SV=2 MTARPPESCVLAKDRPEEPVVPVVSPGPLVPVADFGPLDRLRGWIVTGLITLLATVTRFL NLGSLTDAGTPIFDEKHYAPQAWQVLNNHGVEDNPGYGLVVHPPVGKQLIAIGEAIFGYN GFGWRFTGALLGVVLVALVVRIVRRISRSTLVGAIAGVLLICDGVSFVTARTALLDGFLT FFVVAAFGALIVDRDQVRERMHIALLAGRSAATVWGPRVGVRWWRFGAGVLLGLACATKW SGVYFVLFFGAMALAFDVAARRQYQVQRPWLGTVRRDVLPSGYALGLIPFAVYLATYAPW FASETAIDRHAVGQAVGRNSVVPLPDAVRSLWHYTAKAFHFHAGLTNSAGNYHPWESKPW TWPMSLRPVLYAIDQQDVAGCGAQSCVKAEMLVGTPAMWWLAVPVLAYAGWRMFVRRDWR YAVVLVGYCAGWLPWFADIDRQMYFFYAATMAPFLVMGISLVLGDILYHPGQGSERRTLG LIVVCCYVALVVTNFAWLYPVLTGLPISQQTWNLEIWLPSWR
EMBL CDSCCP43752.1
Sequence length503 AA
Subcellular LocationMembrane
Function in Native Organism
String83332.Rv1002c. 
Potential ApplicationRv1002c-dependent protein O-mannosylation is required for Mtb intracellular persistence and proliferation and for full virulence in SCID mice.
Additional Information1) The attenuation of the mutant virulence is related to the inactivation of Rv1002c gene coding for the Mtb PMT.
2) Rv1002c, a M. tuberculosis membrane protein homolog of eukaryotic protein mannosyltransferases and presented a similar hydropathy profile with PMTs of S. cerevisiae , was shown to catalyze the initial step of protein mannosylation. Bioinformatic analyses shows 11 transmembrane domains in Rv1002c and the presence of the conserved PMT active site residues R39, D55, E56, and R106.
Glycosylation Recation
Glycosylation TypeO- (Ser/Thr) linked 
Mechanism of CatalysisInverting
Basis of catalytic mechnismCatalysis annotation based on experimental evidence.
CAZY FamilyGT39
EC Number (BRENDA)2.4.1.-
Donor TypeLipid linked sugars
Donor SpecificityPolyprenol phosphate -Mannose
Accessory GT IDProGT10.1
Glycan Information
Glycan transferredMonosaccharide (Mannose) 
Method of Glycan IndentificationLiquid chromatography-electrospray ionization-tandem mass spectrometry and affinty chromatography ConA
Genetic evidence of Glycosyltransferase activity
Experimental_strategiesEffect of glycosylation on acceptor protein was analyzed by Gel Electrophoresis and confirmed by mass spectrometry. 
Recombination Host/ Vector usedVector: pSC-B (for knock out) 
Complementation Host/Vector usedvector: pMV361 
Bio Chemical Evidence of Glycosyltransferase Activity
Method of In vitro GT Assay2 mg protein suspended in assay buffer (50 mM MOPS (pH 7.5), 10 mM MgCl2, 1 mM DTT) to which 1.9 nmol [14C] GDP-mannose with a specific activity of 260 mCi/mmol and 1mmol of the synthetic peptide (AAAPPAPATPVAPPPPHHHHHH) were added. The final reaction volume was brought to 500 µL with assay buffer. The assay mixture was incubated at 37°C for 30 min and the enzymatic reaction was stopped by heating at 85°C for 2 min. Suspension centrifuge to pellet particulate material, and the supernatant recovered. Radiolabeled peptides were purified from the supernatant of the enzymatic reaction by Nickel affinity chromatography. Each fraction was added to 10 ml of EcoLume scintillation fluid and counted on a Beckman LS6500 Scintillation counter. 
Recombination Host and vector used in vitro GT assayRv1002c cloned in pMH29 creating pBV90 and electroporated into M. tuberculosis H37Rv and M. smegmatis mc2 155 
Mutantation in recombinant GT1) wild-type Rv1002c and three mutated forms of this gene conferring A55A56, A55E56, or D55A56 substitutions in the invariant D55E56 motif were overexpressed in Mycobacterium smegmatis, and membrane preparations were assayed for in vitro PMT activity. Overexpressed wild-type Mtb Rv1002c significantly increased (66.8%) PMT activity of the M. smegmatis membranes above that observed in the M. smegmatis vector control
2) D55E56 to A55E56 substitution resulted in decreased (56.0%) PMT activity compared with the M. smegmatis vector control. 
Acceptor Subtrate Information
Acceptor Substrate name Alanine and proline-rich secreted protein Apa (50/55-kDa or 45 kDa MPT 32)
ProGPdb ID ProGP51
Experimental Validation of AcceptorIn vitro and in vivo
Organism Mycobacterium tuberculosis H37Rv
Acceptor Substrate name SodC
ProGPdb ID ProGP190
Experimental Validation of AcceptorIn vivo
Litrature
Year Of Validation2005 
Reference Brian C. VanderVen, Jeffery D. Harder, Dean C. Crick, John T. Belisle (2005) Export-mediated assembly of mycobacterial glycoproteins parallels eukaryotic pathways. Science. 2005 Aug 5;309(5736):941-3.

Authors Brian C. VanderVen, Jeffery D. Harder, Dean C. Crick, John T. Belisle
Research groupsMycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA.
Corresponding AuthorJohn T. Belisle
ContactsMycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA.
Reference Liu CF, Tonini L, Malaga W, Beau M, Stella A, Bouyssié D, Jackson MC, Nigou J, Puzo G, Guilhot C, Burlet-Schiltz O, Rivière M(2013). Bacterial protein-O-mannosylating enzyme is crucial for virulence of Mycobacterium tuberculosis.

Authors Liu CF, Tonini L, Malaga W, Beau M, Stella A, Bouyssié D, Jackson MC, Nigou J, Puzo G, Guilhot C, Burlet-Schiltz O, Rivière M
Research groupsScientific Research National Center, Institute of Pharmacology and Structural Biology, F-31077 Toulouse, France.
Corresponding AuthorRivière M.
ContactsScientific Research National Center, Institute of Pharmacology and Structural Biology, F-31077 Toulouse, France.
Reference Córdova-Dávalos LE, Espitia C, González-Cerón G, Arreguín-Espinosa R, Soberón-Chávez G, Servín-González L.(2014). Lipoprotein N -acyl transferase (Lnt1) is dispensable for protein O -mannosylation by Streptomyces coelicolor, 350, 72-82. https://doi.org/10.1111/1574-6968.12298

Authors Córdova-Dávalos LE, Espitia C, González-Cerón G, Arreguín-Espinosa R, Soberón-Chávez G, Servín-González L
Research groupsDepartment of Molecular Biology and Biotechnology, Institute of Biomedical Research, National Autonomous University of Mexico, Ciudad Universitaria, Mexico City, Mexico City.
Corresponding AuthorServín-González L
ContactsDepartment of Molecular Biology and Biotechnology, Institute of Biomedical Research, National Autonomous University of Mexico, Ciudad Universitaria, Mexico City, Mexico City.