Mycobacterium tuberculosis antigens MPT63 and MPT83 increase phagocytic activity of murine peritoneal macrophages

Siromolot, A. A.; Oliinyk, O. S.; Dv, Kolibo; Комісаренко, С. В.

doi:10.15407/ubj88.05.062

Cited by 5 publications

(4 citation statements)

References 17 publications

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“…We employed calcein release assay to investigate if MPT63 would form pores in synthetic membrane. Since MPT63 is known to induce the process of phagocytosis, there could be a plausible correlation between membrane binding, pore formation and phagocytosis. In this assay, the release of calcein dye trapped inside a synthetic vesicle through pores would be monitored by a time-dependent enhancement of fluorescence intensity.…”

Section: Results and Discussionmentioning

confidence: 99%

Conformational Switch Driven Membrane Pore Formation by Mycobacterium Secretory Protein MPT63 Induces Macrophage Cell Death

et al. 2019

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Virulent Mycobacterium tuberculosis (MTB) strains cause cell death of macrophages (Mϕ) inside TB granuloma using a mechanism which is not well understood. Many bacterial systems utilize toxins to induce host cell damage, which occurs along with immune evasion. These toxins often use chameleon sequences to generate an environmentsensitive conformational switch, facilitating the process of infection. The presence of toxins is not yet known for MTB. Here, we show that MTBsecreted immunogenic MPT63 protein undergoes a switch from β-sheet to helix in response to mutational and environmental stresses. MPT63 in its helical form creates pores in both synthetic and Mϕ membranes, while the native β-sheet protein remains inert toward membrane interactions. Using fluorescence correlation spectroscopy and atomic force microscopy, we show further that the helical form undergoes self-association to produce toxic oligomers of different morphology. Trypan blue and flow cytometry analyses reveal that the helical state can be utilized by MTB for killing Mϕ cells. Collectively, our study emphasizes for the first time a toxin-like behavior of MPT63 induced by an environment-dependent conformational switch, resulting in membrane pore formation by toxic oligomers and Mϕ cell death.

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Section: Results and Discussionmentioning

confidence: 99%

Conformational Switch Driven Membrane Pore Formation by Mycobacterium Secretory Protein MPT63 Induces Macrophage Cell Death

et al. 2019

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“…MPT63 was characterized by SDS-PAGE and measuring 16 kDa (Figure 5). MPT63 with a molecular weight of 16 kDa is a secreted protein that has a high level of expression in mycobacteria [14].…”

Section: Resultsmentioning

confidence: 99%

“…E. coli BL21 carrying a recombinant plasmid pQE-Rv1926c Lysis of recombinant clone cells was carried out by sonication to obtain MPT63 recombinant protein.MPT63 was characterized by SDS-PAGE and measuring 16 kDa (Figure5). MPT63 with a molecular weight of 16 kDa is a secreted protein that has a high level of expression in mycobacteria[14].…”

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confidence: 99%

Cloning and Expression of Recombinant Protein MPT63 of Mycobacterium tuberculosis Indonesian Isolate as Serodiagnostic Latent Tuberculosis

Agus¹,

Fidhatami²,

Hatta³

2021

Proceedings of the 3rd KOBI Congress, International and National Conferences (KOBICINC 2020)

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Indonesia and 13 other countries are included in High Burden Countries (HBC) in Tuberculosis (TB) cases. One of the problems in TB control in Indonesia is detecting latent TB appropriately. Diagnosis of latent TB does not have a gold standard. Some tests to detect latent TB have limitations in TB endemic areas such as Indonesia. Therefore, we need a fast, effective, and accurate method to diagnose TB. One of the potential proteins is MPT63, which is coded by the Rv 1926c gene. It is known that MPT 63 can induce Th1 cell reactivity and proliferation of IFN-λ. The purpose of this research was to clone and produce the recombinant protein MPT 63 from Mycobacterium tuberculosis. The method is Rv 1926c ligation to the pQE30-Xa expression vector and transformation to Escherichia coli BL21 host cell. Recombinant protein production was carried out by growing recombinant clones Rv1926-pQE on Luria Bertani medium by IPTG induction. The transformation results obtained white colonies and blue colonies. The growth of Escherichia coli BL21 carrying a recombinant plasmid is characterized by a change in the color of the medium. MPT63 recombinant protein was characterized by SDS-PAGE with a size of 16 kDa.

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“…In previous studies, MPT63 induced cell death of macrophages through a conformational switch dependent on the pH of the host [ 8 ]. Additionally, MPT63 enhanced the phagocytic activity by upregulating the secretion of TNF-α and IL-6 in murine peritoneal macrophages [ 9 ]. Located in the RD2 region, MPT64 induced the production of IFN-γ in murine macrophages and patients with TB, and reduced apoptosis by increasing the expression of TGF-β and reducing inflammation [ 10 , 11 ].…”

Section: Introductionmentioning

confidence: 99%

Multi-Functional MPT Protein as a Therapeutic Agent against Mycobacterium tuberculosis

et al. 2021

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Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis (TB), avoids the host immune system through its virulence factors. MPT63 and MPT64 are the virulence factors secreted by MTB which regulate host proteins for the survival and proliferation of MTB in the host. Here, we found that MPT63 bound directly with TBK1 and p47phox, whereas MPT64 interacted with TBK1 and HK2. We constructed a MPT63/64-derived multifunctional recombinant protein (rMPT) that was able to interact with TBK1, p47phox, or HK2. rMPT was shown to regulate IFN-β levels and increase inflammation and concentration of reactive oxygen species (ROS), while targeting macrophages and killing MTB, both in vitro and in vivo. Furthermore, the identification of the role of rMPT against MTB was achieved via vaccination in a mouse model. Taken together, we here present rMPT, which, by regulating important immune signaling systems, can be considered an effective vaccine or therapeutic agent against MTB.

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Mycobacterium tuberculosis antigens MPT63 and MPT83 increase phagocytic activity of murine peritoneal macrophages

Cited by 5 publications

References 17 publications

Conformational Switch Driven Membrane Pore Formation by Mycobacterium Secretory Protein MPT63 Induces Macrophage Cell Death

Conformational Switch Driven Membrane Pore Formation by Mycobacterium Secretory Protein MPT63 Induces Macrophage Cell Death

Cloning and Expression of Recombinant Protein MPT63 of Mycobacterium tuberculosis Indonesian Isolate as Serodiagnostic Latent Tuberculosis

Multi-Functional MPT Protein as a Therapeutic Agent against Mycobacterium tuberculosis

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