Structures of monomeric and oligomeric forms of the
            <i>Toxoplasma gondii</i>
            perforin-like protein 1

Tien, Ni; Williams, Sophie I.; Rezelj, Saša; Anderluh, Gregor; Harlos, K.; Stansfeld, Phillip J.; Gilbert, Robert J.C.

doi:10.1126/sciadv.aaq0762

Cited by 31 publications

(42 citation statements)

References 55 publications

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“…As observed in other MACPF/CDC proteins, such as the Apicomplexan perforin-like protein TgPLP1 (17) and the pneumolysin prepore (18,19), the MACPF domains make extensive complementary contacts with each other in the PFN2 pre-pore structure (Fig. 1H).…”

Section: Cryo-em Structure Of the Pfn2 Pre-pore Oligomersupporting

confidence: 62%

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Structure and mechanism of bactericidal mammalian perforin-2, an ancient agent of innate immunity

et al. 2020

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Perforin-2 (MPEG1) is thought to enable the killing of invading microbes engulfed by macrophages and other phagocytes, forming pores in their membranes. Loss of perforin-2 renders individual phagocytes and whole organisms significantly more susceptible to bacterial pathogens. Here, we reveal the mechanism of perforin-2 activation and activity using atomic structures of pre-pore and pore assemblies, high-speed atomic force microscopy, and functional assays. Perforin-2 forms a pre-pore assembly in which its pore-forming domain points in the opposite direction to its membrane-targeting domain. Acidification then triggers pore formation, via a 180° conformational change. This novel and unexpected mechanism prevents premature bactericidal attack and may have played a key role in the evolution of all perforin family proteins.

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Section: Cryo-em Structure Of the Pfn2 Pre-pore Oligomersupporting

confidence: 62%

“…S3, and table S2). The P2 domain has pseudo-threefold symmetry ( Apicomplexan perforin-like proteins (17). Each repeat contains three strands with a conserved interstrand disulfide bond [pairwise root mean square deviation (RMSD) = 0.76 to 1.00 Å calculated from the three repeats].…”

Section: The  Hairpin Of the P2 Domain Directs Membrane Recognitionmentioning

confidence: 99%

Structure and mechanism of bactericidal mammalian perforin-2, an ancient agent of innate immunity

et al. 2020

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“…PPLP molecules are initially secreted as monomers which bind to the target cell membrane and oligomerize on its surface to form functional, transmembrane pores(18,21). The structure of PPLPs remain unresolved, however, recently the crystal structure of a closely related PLP of another apicomplexan parasite Toxoplasma gondii , TgPLP1 was deciphered (22). The structure of TgPLP1 is similar to the reported eukaryotic pore forming proteins.…”

Section: Introductionmentioning

confidence: 99%

“…The structure of TgPLP1 is similar to the reported eukaryotic pore forming proteins. But, it is bulkier than others in terms of the presence of extra helices that may play roles in pore formation (22). The structure of PPLPs can now be predicted using TgPLP1 as template for designing anti-malarial chemotherapeutics.…”

Section: Introductionmentioning

confidence: 99%

Supression of Perforin-like Protein pores inhibitPlasmodiummultistage-growth, transmission and erythrocyte senescence

Garg

Shivappagowdar

et al. 2019

Preprint

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25The pore forming Plasmodium perforin like proteins (PPLP), expressed in all stages of 26 the parasite life cycle are central drivers for host interactions critical for completion of 27 parasite life cycle and high transmission rates. The high sequence similarity in the central 28 membrane attack complex/ perforin (MACPF) domain and consequent functional overlaps 29 defines them as an attractive target for the development of multi-stage antimalarials. Herein 30 we evaluated the mechanism of pan active function of central, highly conserved region of 31 PPLPs, MACPF domain (PMD) and inhibitory potential of specifically designed anti-PMD 32 chemo. The E. coli expressed rPMD interacts with erythrocyte membrane and form pores of 33 ~10.5 nm height and ~24.3 nm diameter leading to haemoglobin release and dextran uptake. 34The treatment with PMD induced erythrocytes senescence at 48 hours which can account for 35 the physiological effect of disseminated PLPs in loss of circulating erythrocytes inducing 36 anemia during malaria infection. The anti-PMD inhibitors effectively blocked 37 intraerythrocytic growth by suppressing invasion and egress of merozoites and protecting 38 against erythrocyte senescence. Moreover, these inhibitors also blocked the hepatic stage and 39 transmission stage parasite development suggesting multi-stage and transmission-blocking 40 potential of these inhibitors. Additionally, the erythrocyte senescence protective potential of 41 PMD inhibitors can be used to occlude PPLPs mediated severe malarial anemia. Further these 42 inhibitors can be developed with a potential to protect against severity of the disease. 43 44 45 Author Summary 46Malaria continues to be a major global health threat despite of several exciting improvements 47 in the treatment and prevention of the disease. One of the major concerns in the development 48 of therapy is the emergence of the drug resistance. But for the efficient treatment regime, 49 targeting multiple stages including host and vector would serve as an ideal therapy. Perforin 50 like proteins (PLPs) are eukaryotic pore forming proteins that are highly conserved in the 51 apicomplexan parasites. These play crucial roles in entry and exit of parasites from the host 52 cells and establish infection at multiple stages of Plasmodium spp. life cycle. Understanding 53 the mechanism of pore formation by smaller, functional, pan-active scaffold of PLPs can 54 serve as a target for development of cross stage protection. Here, using various biochemical, 55 biophysical and pharmacological evidences, we validate the activity and characterize the pore 56 formation of PLPs on erythrocytes. Further, our specifically designed inhibitors could restrict 57 this pore formation and impede the exit/entry of the parasites. Moreover, these inhibitors 58 could also exert multiple stage inhibition and rescue the uninfected erythrocytes from death. 59Together, this study highlights the mechanism of pore formation by PPLPs and evaluates 60 their potential for the development of...

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“…The final pore comprises a large (compete or incomplete) β-barrel that spans the membrane (19,23). In contrast to other MACPF/CDCs studied to date (19,(24)(25)(26)(27), the recombinant form of MPEG-1 has pre-assembled into a soluble oligomer that exhibits the expected features of a pre-pore assembly. Within each subunit, both TMH-1 and -2 are folded as small helical bundles (Fig.…”

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confidence: 99%

The structure of Macrophage Expressed Gene-1, a phagolysosome immune effector that is activated upon acidification

Pang

Bayly-Jones

Radjainia

et al. 2019

Preprint

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Macrophage Expressed Gene-1 (MPEG-1; also termed Perforin-2) is an endosomal / phagolysosomal perforin-like protein that is conserved across the metazoan kingdom and that functions within the phagolysosome to damage engulfed microbes. Like the Membrane Attack Complex and perforin, MPEG-1 has been postulated to form pores in target membranes, however, its mode of action remains to be established. We used single particle cryo-Electron Microscopy to determine the 2.4 Å structure of a hexadecameric assembly of MPEG-1 that displays the expected features of a soluble pre-pore complex. We further discovered that the MPEG-1 pre-pore-like assemblies can be induced to perforate membranes through mild acidification, such as would occur within maturing phagolysosomes. We next solved the 3.6 Å cryo-EM structure of MPEG-1 in complex with liposomes. Remarkably these data revealed that a C-terminal Multi-vesicular body of 12 kDa (MVB12)-associatedβ-prism (MABP) domain interacts with target membranes in a mode that positions the pore forming machinery of MPEG-1 to point away from the bound membrane. This unexpected mechanism of membrane interaction raises the intriguing possibility that MPEG-1 may be able to remain bound to the phagolysosome membrane while simultaneously forming pores in engulfed bacterial targets.

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Structures of monomeric and oligomeric forms of the Toxoplasma gondii perforin-like protein 1

Abstract: Structures of Toxoplasma perforin-like protein facilitate understanding of the protein’s role in infection and disease.

Cited by 31 publications

References 55 publications

Structure and mechanism of bactericidal mammalian perforin-2, an ancient agent of innate immunity

Structure and mechanism of bactericidal mammalian perforin-2, an ancient agent of innate immunity

Supression of Perforin-like Protein pores inhibitPlasmodiummultistage-growth, transmission and erythrocyte senescence

The structure of Macrophage Expressed Gene-1, a phagolysosome immune effector that is activated upon acidification

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