Structural basis of Toxoplasma gondii perforin-like protein 1 membrane interaction and activity during egress

Guerra, Alfredo J.; Zhang, Ou; Bahr, Constance m. E.; Huynh, My-Hang; DelProposto, James; Brown, William C.; Wawrzak, Zdzislaw; Koropatkin, Nicole M.; Carruthers, Vern B.

doi:10.1371/journal.ppat.1007476

Title: Structural basis of Toxoplasma gondii perforin-like protein 1 membrane interaction and activity during egress

Journal Article · Tue Dec 04 00:00:00 EST 2018 · PLoS Pathogens

DOI:https://doi.org/10.1371/journal.ppat.1007476· OSTI ID:1491870

^[1];

^[1]; Bahr, Constance m. E. ^[1]; Huynh, My-Hang ^[1]; DelProposto, James ^[1]; Brown, William C. ^[1]; Wawrzak, Zdzislaw ^[2];

^[1]; Carruthers, Vern B. ^[1]

Univ. of Michigan, Ann Arbor, MI (United States)
Northwestern Univ., Argonne, IL (United States)

Intracellular pathogens must egress from the host cell to continue their infectious cycle. Apicomplexans are a phylum of intracellular protozoans that have evolved members of the membrane attack complex and perforin (MACPF) family of pore forming proteins to disrupt cellular membranes for traversing cells during tissue migration or egress from a replicative vacuole following intracellular reproduction. Previous work showed that the apicomplexan Toxoplasma gondii secretes a perforin-like protein (TgPLP1) that contains a C-terminal Domain (CTD) which is necessary for efficient parasite egress. However, the structural basis for CTD membrane binding and egress competency remained unknown. Here, we present evidence that TgPLP1 CTD prefers binding lipids that are abundant in the inner leaflet of the lipid bilayer. Additionally, solving the high-resolution crystal structure of the TgPLP1 APCβ domain within the CTD reveals an unusual double-layered β-prism fold that resembles only one other protein of known structure. Three direct repeat sequences comprise subdomains, with each constituting a wall of the β-prism fold. One subdomain features a protruding hydrophobic loop with an exposed tryptophan at its tip. Spectrophotometric measurements of intrinsic tryptophan fluorescence are consistent with insertion of the hydrophobic loop into a target membrane. Using CRISPR/Cas9 gene editing we show that parasite strains bearing mutations in the hydrophobic loop, including alanine substitution of the tip tryptophan, are equally deficient in egress as a strain lacking TgPLP1 altogether. Taken together our findings suggest a crucial role for the hydrophobic loop in anchoring TgPLP1 to the membrane to support its cytolytic activity and egress function.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-06CH11357; R01AI46675

OSTI ID:: 1491870

Journal Information:: PLoS Pathogens, Vol. 14, Issue 12; ISSN 1553-7374

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

Citation Metrics:

Cited by: 9 works

Citation information provided by
Web of Science

References (48)

Signalling mechanisms involved in apical organelle discharge during host cell invasion by apicomplexan parasites Singh, Shailja; Chitnis, Chetan E. Microbes and Infection, Vol. 14, Issue 10 https://doi.org/10.1016/j.micinf.2012.05.007	journal	August 2012
The moving junction of apicomplexan parasites: a key structure for invasion: The moving junction of apicomplexan parasites Besteiro, Sébastien; Dubremetz, Jean-François; Lebrun, Maryse Cellular Microbiology, Vol. 13, Issue 6 https://doi.org/10.1111/j.1462-5822.2011.01597.x	journal	April 2011
Phagosome Extrusion and Host-Cell Survival after Cryptococcus neoformans Phagocytosis by Macrophages Alvarez, Mauricio; Casadevall, Arturo Current Biology, Vol. 16, Issue 21 https://doi.org/10.1016/j.cub.2006.09.061	journal	November 2006
A cytolytic protein from the edible mushroom, Pleurotus ostreatus Bernheimer, Alan W.; Avigad, Lois S. Biochimica et Biophysica Acta (BBA) - General Subjects, Vol. 585, Issue 3 https://doi.org/10.1016/0304-4165(79)90090-4	journal	July 1979
Structural Basis for Receptor Recognition by the Human CD59-Responsive Cholesterol-Dependent Cytolysins Lawrence, Sara L.; Gorman, Michael A.; Feil, Susanne C. Structure, Vol. 24, Issue 9 https://doi.org/10.1016/j.str.2016.06.017	journal	September 2016
Structures of Lysenin Reveal a Shared Evolutionary Origin for Pore-Forming Proteins And Its Mode of Sphingomyelin Recognition De Colibus, Luigi; Sonnen, Andreas F. -P.; Morris, Keith J. Structure, Vol. 20, Issue 9 https://doi.org/10.1016/j.str.2012.06.011	journal	September 2012
Packing a punch: the mechanism of pore formation by cholesterol dependent cytolysins and membrane attack complex/perforin-like proteins Dunstone, Michelle A.; Tweten, Rodney K. Current Opinion in Structural Biology, Vol. 22, Issue 3 https://doi.org/10.1016/j.sbi.2012.04.008	journal	June 2012
Evidence for Host Cells as the Major Contributor of Lipids in the Intravacuolar Network of Toxoplasma-Infected Cells Caffaro, Carolina E.; Boothroyd, John C. Eukaryotic Cell, Vol. 10, Issue 8 https://doi.org/10.1128/EC.00002-11	journal	June 2011
Reverse genetics screen identifies six proteins important for malaria development in the mosquito: Proteins important for malaria development in the mosquito Ecker, Andrea; Bushell, Ellen S. C.; Tewari, Rita Molecular Microbiology, Vol. 70, Issue 1 https://doi.org/10.1111/j.1365-2958.2008.06407.x	journal	August 2008
Actin-dependent movement of bacterial pathogens Stevens, Joanne M.; Galyov, Edouard E.; Stevens, Mark P. Nature Reviews Microbiology, Vol. 4, Issue 2 https://doi.org/10.1038/nrmicro1320	journal	February 2006
Dali server update Holm, Liisa; Laakso, Laura M. Nucleic Acids Research, Vol. 44, Issue W1 https://doi.org/10.1093/nar/gkw357	journal	April 2016
Challenges in the successful control of the avian coccidia Shirley, Martin W.; Smith, Adrian L.; Blake, Damer P. Vaccine, Vol. 25, Issue 30 https://doi.org/10.1016/j.vaccine.2006.12.030	journal	July 2007
Structural elements of the cholesterol-dependent cytolysins that are responsible for their cholesterol-sensitive membrane interactions Soltani, C. E.; Hotze, E. M.; Johnson, A. E. Proceedings of the National Academy of Sciences, Vol. 104, Issue 51 https://doi.org/10.1073/pnas.0708104105	journal	December 2007
Cell Traversal Activity Is Important for Plasmodium falciparum Liver Infection in Humanized Mice Yang, Annie S. P.; O’Neill, Matthew T.; Jennison, Charlie Cell Reports, Vol. 18, Issue 13 https://doi.org/10.1016/j.celrep.2017.03.017	journal	March 2017
Toxoplasma gondii: 25 years and 25 major advances for the field Boothroyd, John C. International Journal for Parasitology, Vol. 39, Issue 8 https://doi.org/10.1016/j.ijpara.2009.02.003	journal	July 2009
Lipid Organization of the Plasma Membrane Ingólfsson, Helgi I.; Melo, Manuel N.; van Eerden, Floris J. Journal of the American Chemical Society, Vol. 136, Issue 41 https://doi.org/10.1021/ja507832e	journal	October 2014
CryoEM structures of membrane pore and prepore complex reveal cytolytic mechanism of Pneumolysin van Pee, Katharina; Neuhaus, Alexander; D'Imprima, Edoardo eLife, Vol. 6 https://doi.org/10.7554/eLife.23644	journal	March 2017
Host Cell Traversal Is Important for Progression of the Malaria Parasite through the Dermis to the Liver Amino, Rogerio; Giovannini, Donatella; Thiberge, Sabine Cell Host & Microbe, Vol. 3, Issue 2 https://doi.org/10.1016/j.chom.2007.12.007	journal	February 2008
From The Cover: Essential role of membrane-attack protein in malarial transmission to mosquito host Kadota, K.; Ishino, T.; Matsuyama, T. Proceedings of the National Academy of Sciences, Vol. 101, Issue 46 https://doi.org/10.1073/pnas.0406187101	journal	November 2004
PHENIX: a comprehensive Python-based system for macromolecular structure solution Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221 https://doi.org/10.1107/S0907444909052925	journal	January 2010
The glideosome: a molecular machine powering motility and host-cell invasion by Apicomplexa Keeley, Anthony; Soldati, Dominique Trends in Cell Biology, Vol. 14, Issue 10 https://doi.org/10.1016/j.tcb.2004.08.002	journal	October 2004
Mechanisms of host cell exit by the intracellular bacterium Chlamydia Hybiske, K.; Stephens, R. S. Proceedings of the National Academy of Sciences, Vol. 104, Issue 27 https://doi.org/10.1073/pnas.0703218104	journal	June 2007
Lysenin, a Novel Sphingomyelin-specific Binding Protein Yamaji, Akiko; Sekizawa, Yoshiyuki; Emoto, Kazuo Journal of Biological Chemistry, Vol. 273, Issue 9 https://doi.org/10.1074/jbc.273.9.5300	journal	February 1998
Mechanism of Membrane Insertion of a Multimeric β-Barrel Protein Heuck, Alejandro P.; Hotze, Eileen M.; Tweten, Rodney K. Molecular Cell, Vol. 6, Issue 5 https://doi.org/10.1016/S1097-2765(00)00119-2	journal	November 2000
Functional Dissection of Toxoplasma gondii Perforin-like Protein 1 Reveals a Dual Domain Mode of Membrane Binding for Cytolysis and Parasite Egress Roiko, Marijo S.; Carruthers, Vern B. Journal of Biological Chemistry, Vol. 288, Issue 12 https://doi.org/10.1074/jbc.M113.450932	journal	February 2013
Rapid Membrane Disruption by a Perforin-Like Protein Facilitates Parasite Exit from Host Cells Kafsack, B. F. C.; Pena, J. D. O.; Coppens, I. Science, Vol. 323, Issue 5913 https://doi.org/10.1126/science.1165740	journal	January 2009
A Plasmodium sporozoite protein with a membrane attack complex domain is required for breaching the liver sinusoidal cell layer prior to hepatocyte infection†: Role of a MACPF-related protein in the malarial life cycle Ishino, Tomoko; Chinzei, Yasuo; Yuda, Masao Cellular Microbiology, Vol. 7, Issue 2 https://doi.org/10.1111/j.1462-5822.2004.00447.x	journal	September 2004
A Pore-forming Toxin Interacts with a GPI-anchored Protein and Causes Vacuolation of the Endoplasmic Reticulum Abrami, Laurence; Fivaz, Marc; Glauser, Pierre-Etienne The Journal of Cell Biology, Vol. 140, Issue 3 https://doi.org/10.1083/jcb.140.3.525	journal	February 1998
Structural Basis for Recognition of the Pore-Forming Toxin Intermedilysin by Human Complement Receptor CD59 Johnson, Steven; Brooks, Nicholas J.; Smith, Richard A. G. Cell Reports, Vol. 3, Issue 5 https://doi.org/10.1016/j.celrep.2013.04.029	journal	May 2013
Perforin-like protein PPLP4 is crucial for mosquito midgut infection by Plasmodium falciparum Wirth, Christine C.; Bennink, Sandra; Scheuermayer, Matthias Molecular and Biochemical Parasitology, Vol. 201, Issue 2 https://doi.org/10.1016/j.molbiopara.2015.06.005	journal	June 2015
Data processing and analysis with the autoPROC toolbox Vonrhein, Clemens; Flensburg, Claus; Keller, Peter Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4 https://doi.org/10.1107/S0907444911007773	journal	March 2011
Interaction of .theta.-Toxin (Perfringolysin O), a Cholesterol-Binding Cytolysin, with Liposomal Membranes: Change in the Aromatic Side Chains upon Binding and Insertion Nakamura, Megumi; Sekino, Naoko; Iwamoto, Machiko Biochemistry, Vol. 34, Issue 19 https://doi.org/10.1021/bi00019a032	journal	May 1995
Molecular mechanisms of host cell traversal by malaria sporozoites Yang, Annie S. P.; Boddey, Justin A. International Journal for Parasitology, Vol. 47, Issue 2-3 https://doi.org/10.1016/j.ijpara.2016.09.002	journal	February 2017
Expulsion of Live Pathogenic Yeast by Macrophages Ma, Hansong; Croudace, Joanne E.; Lammas, David A. Current Biology, Vol. 16, Issue 21 https://doi.org/10.1016/j.cub.2006.09.032	journal	November 2006
Cryptosporidiosis: host immune responses and the prospects for effective immunotherapies McDonald, Vincent Expert Review of Anti-infective Therapy, Vol. 9, Issue 11 https://doi.org/10.1586/eri.11.123	journal	November 2011
Membrane assembly of the cholesterol-dependent cytolysin pore complex Hotze, Eileen M.; Tweten, Rodney K. Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1818, Issue 4 https://doi.org/10.1016/j.bbamem.2011.07.036	journal	April 2012
Pleurotolysin, a Novel Sphingomyelin-specific Two-component Cytolysin from the Edible Mushroom Pleurotus ostreatus , Assembles into a Transmembrane Pore Complex Tomita, Toshio; Noguchi, Kayoko; Mimuro, Hitomi Journal of Biological Chemistry, Vol. 279, Issue 26 https://doi.org/10.1074/jbc.M402676200	journal	April 2004
Perforin-like protein PPLP2 permeabilizes the red blood cell membrane during egress of P lasmodium falciparum gametocytes : PPLP2 in erythrocyte egress by malaria gametocytes Wirth, Christine C.; Glushakova, Svetlana; Scheuermayer, Matthias Cellular Microbiology, Vol. 16, Issue 5 https://doi.org/10.1111/cmi.12288	journal	April 2014
Toxoplasma Parasite Twisting Motion Mechanically Induces Host Cell Membrane Fission to Complete Invasion within a Protective Vacuole Pavlou, Georgios; Biesaga, Mateusz; Touquet, Bastien Cell Host & Microbe, Vol. 24, Issue 1 https://doi.org/10.1016/j.chom.2018.06.003	journal	July 2018
Membrane cholesterol and sphingomyelin, and ostreolysin A are obligatory for pore-formation by a MACPF/CDC-like pore-forming protein, pleurotolysin B Ota, Katja; Leonardi, Adrijana; Mikelj, Miha Biochimie, Vol. 95, Issue 10 https://doi.org/10.1016/j.biochi.2013.06.012	journal	October 2013
Human CD59 is a receptor for the cholesterol-dependent cytolysin intermedilysin Giddings, Kara S.; Zhao, Ji; Sims, Peter J. Nature Structural & Molecular Biology, Vol. 11, Issue 12 https://doi.org/10.1038/nsmb862	journal	November 2004
Binding of perforin to membranes is sensitive to lipid spacing and not headgroup Antia, R.; Schlegel, R.; Williamson, P. Immunology Letters, Vol. 32, Issue 2 https://doi.org/10.1016/0165-2478(92)90108-z	journal	April 1992
The structural basis for membrane binding and pore formation by lymphocyte perforin Law, Ruby H. P.; Lukoyanova, Natalya; Voskoboinik, Ilia Nature, Vol. 468, Issue 7322 https://doi.org/10.1038/nature09518	journal	October 2010
The Obligate Intracellular Parasite Toxoplasma gondii Secretes a Soluble Phosphatidylserine Decarboxylase Gupta, Nishith; Hartmann, Anne; Lucius, Richard Journal of Biological Chemistry, Vol. 287, Issue 27 https://doi.org/10.1074/jbc.m112.373639	journal	May 2012
Molecular Determinants of Sphingomyelin Specificity of a Eukaryotic Pore-forming Toxin Bakrač, Biserka; Gutiérrez-Aguirre, Ion; Podlesek, Zdravko Journal of Biological Chemistry, Vol. 283, Issue 27 https://doi.org/10.1074/jbc.m708747200	journal	April 2008
A Conserved Apicomplexan Microneme Protein Contributes to Toxoplasma gondii Invasion and Virulence Huynh, My-Hang; Boulanger, Martin J.; Carruthers, Vern B. Infection and Immunity, Vol. 82, Issue 10 https://doi.org/10.1128/iai.01877-14	journal	August 2014
Apoptosis, Pyroptosis, and Necrosis: Mechanistic Description of Dead and Dying Eukaryotic Cells Fink, S. L.; Cookson, B. T. Infection and Immunity, Vol. 73, Issue 4 https://doi.org/10.1128/iai.73.4.1907-1916.2005	journal	March 2005
The structural basis for membrane binding and pore formation by lymphocyte perforin Law, Ruby; Lukoyanova, Natalya; Voskoboinik, Ilia Monash University https://doi.org/10.4225/03/57428c5b7f179	dataset	January 2016

Cited By (1)

mSphere of Influence: Ushering in the CRISPR Revolution to Toxoplasma Biology Guerra, Alfredo J. mSphere, Vol. 4, Issue 4 https://doi.org/10.1128/msphere.00307-19	journal	August 2019

Similar Records

Structural and Functional Divergence of the Aldolase Fold in Toxoplasma gondii

Journal Article · Thu Oct 02 00:00:00 EDT 2014 · Journal of Molecular Biology · OSTI ID:1491870

Tonkin, Michelle L.; Halavaty, Andrei S.; Ramaswamy, Raghavendran; +4 more

Toxoplasma gondii: Biochemical and biophysical characterization of recombinant soluble dense granule proteins GRA2 and GRA6

Journal Article · Fri Mar 27 00:00:00 EDT 2015 · Biochemical and Biophysical Research Communications · OSTI ID:1491870

Bittame, Amina; Effantin, Grégory; Pètre, Graciane; +7 more

Studies of Toxoplasma gondii and Plasmodium falciparum enoyl acyl carrier protein reductase and implications for the development of antiparasitic agents

Journal Article · Thu Mar 01 00:00:00 EST 2007 · Acta Crystallographica. Section D: Biological Crystallography · OSTI ID:1491870

Muench, Stephen P.; Prigge, Sean T.; McLeod, Rima; +4 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES

Title: Structural basis of Toxoplasma gondii perforin-like protein 1 membrane interaction and activity during egress

Citation Formats

References (48)

Cited By (1)

Similar Records

Related Subjects