Biophysics of Malarial Parasite Exit from Infected Erythrocytes
Upon infection and development within human erythrocytes, P. falciparum induces alterations to the infected RBC morphology and bio-mechanical properties to eventually rupture the host cells through parasitic and host derived proteases of cysteine and serine families. We used previously reported broa...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Public Library of Science
2011
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| Online Access: | http://hdl.handle.net/1721.1/65418 https://orcid.org/0000-0002-6223-6831 |
| _version_ | 1826216747180490752 |
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| author | Chandramohanadas, Rajesh Park, YongKeun Lui, Lena Li, Ang Quinn, David Liew, Kingsley Diez Silva, Monica Sung, Yong-Jin Dao, Ming Lim, Chwee Teck Preiser, Peter Rainer Suresh, Subra |
| author2 | Massachusetts Institute of Technology. Spectroscopy Laboratory |
| author_facet | Massachusetts Institute of Technology. Spectroscopy Laboratory Chandramohanadas, Rajesh Park, YongKeun Lui, Lena Li, Ang Quinn, David Liew, Kingsley Diez Silva, Monica Sung, Yong-Jin Dao, Ming Lim, Chwee Teck Preiser, Peter Rainer Suresh, Subra |
| author_sort | Chandramohanadas, Rajesh |
| collection | MIT |
| description | Upon infection and development within human erythrocytes, P. falciparum induces alterations to the infected RBC morphology and bio-mechanical properties to eventually rupture the host cells through parasitic and host derived proteases of cysteine and serine families. We used previously reported broad-spectrum inhibitors (E64d, EGTA-AM and chymostatin) to inhibit these proteases and impede rupture to analyze mechanical signatures associated with parasite escape. Treatment of late-stage iRBCs with E64d and EGTA-AM prevented rupture, resulted in no major RBC cytoskeletal reconfiguration but altered schizont morphology followed by dramatic re-distribution of three-dimensional refractive index (3D-RI) within the iRBC. These phenotypes demonstrated several-fold increased iRBC membrane flickering. In contrast, chymostatin treatment showed no 3D-RI changes and caused elevated fluctuations solely within the parasitophorous vacuole. We show that E64d and EGTA-AM supported PV breakdown and the resulting elevated fluctuations followed non-Gaussian pattern that resulted from direct merozoite impingement against the iRBC membrane. Optical trapping experiments highlighted reduced deformability of the iRBC membranes upon rupture-arrest, more specifically in the treatments that facilitated PV breakdown. Taken together, our experiments provide novel mechanistic interpretations on the role of parasitophorous vacuole in maintaining the spherical schizont morphology, the impact of PV breakdown on iRBC membrane fluctuations leading to eventual parasite escape and the evolution of membrane stiffness properties of host cells in which merozoites were irreversibly trapped, recourse to protease inhibitors. These findings provide a comprehensive, previously unavailable, body of information on the combined effects of biochemical and biophysical factors on parasite egress from iRBCs. |
| first_indexed | 2024-09-23T16:52:46Z |
| format | Article |
| id | mit-1721.1/65418 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:52:46Z |
| publishDate | 2011 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/654182022-10-03T08:52:18Z Biophysics of Malarial Parasite Exit from Infected Erythrocytes Chandramohanadas, Rajesh Park, YongKeun Lui, Lena Li, Ang Quinn, David Liew, Kingsley Diez Silva, Monica Sung, Yong-Jin Dao, Ming Lim, Chwee Teck Preiser, Peter Rainer Suresh, Subra Massachusetts Institute of Technology. Spectroscopy Laboratory Suresh, Subra Suresh, Subra Chandramohanadas, Rajesh Park, YongKeun Lui, Lena Li, Ang Quinn, David Liew, Kingsley Diez Silva, Monica Sung, Yongjin Dao, Ming Upon infection and development within human erythrocytes, P. falciparum induces alterations to the infected RBC morphology and bio-mechanical properties to eventually rupture the host cells through parasitic and host derived proteases of cysteine and serine families. We used previously reported broad-spectrum inhibitors (E64d, EGTA-AM and chymostatin) to inhibit these proteases and impede rupture to analyze mechanical signatures associated with parasite escape. Treatment of late-stage iRBCs with E64d and EGTA-AM prevented rupture, resulted in no major RBC cytoskeletal reconfiguration but altered schizont morphology followed by dramatic re-distribution of three-dimensional refractive index (3D-RI) within the iRBC. These phenotypes demonstrated several-fold increased iRBC membrane flickering. In contrast, chymostatin treatment showed no 3D-RI changes and caused elevated fluctuations solely within the parasitophorous vacuole. We show that E64d and EGTA-AM supported PV breakdown and the resulting elevated fluctuations followed non-Gaussian pattern that resulted from direct merozoite impingement against the iRBC membrane. Optical trapping experiments highlighted reduced deformability of the iRBC membranes upon rupture-arrest, more specifically in the treatments that facilitated PV breakdown. Taken together, our experiments provide novel mechanistic interpretations on the role of parasitophorous vacuole in maintaining the spherical schizont morphology, the impact of PV breakdown on iRBC membrane fluctuations leading to eventual parasite escape and the evolution of membrane stiffness properties of host cells in which merozoites were irreversibly trapped, recourse to protease inhibitors. These findings provide a comprehensive, previously unavailable, body of information on the combined effects of biochemical and biophysical factors on parasite egress from iRBCs. Singapore. Agency for Science, Technology and Research Singapore-MIT Alliance Global Enterprise for Micro-Mechanics and Molecular Medicine National University of Singapore National Institutes of Health (U.S.) (Grant R01 HL094270-01A1) National Institutes of Health (U.S.) (Grant 1-R01-GM076689-01) National Institutes of Health (U.S.) (P41-RR02594-18-24) 2011-08-26T19:46:55Z 2011-08-26T19:46:55Z 2011-06 2011-02 Article http://purl.org/eprint/type/JournalArticle 1932-6203 http://hdl.handle.net/1721.1/65418 Chandramohanadas, Rajesh et al. “Biophysics of Malarial Parasite Exit from Infected Erythrocytes.” Ed. Volker Theo Heussler. PLoS ONE 6.6 (2011) : e20869. https://orcid.org/0000-0002-6223-6831 en_US http://dx.doi.org/10.1371/journal.pone.0020869 PLoS ONE Creative Commons Attribution http://creativecommons.org/licenses/by/2.5/ application/pdf Public Library of Science PLoS |
| spellingShingle | Chandramohanadas, Rajesh Park, YongKeun Lui, Lena Li, Ang Quinn, David Liew, Kingsley Diez Silva, Monica Sung, Yong-Jin Dao, Ming Lim, Chwee Teck Preiser, Peter Rainer Suresh, Subra Biophysics of Malarial Parasite Exit from Infected Erythrocytes |
| title | Biophysics of Malarial Parasite Exit from Infected Erythrocytes |
| title_full | Biophysics of Malarial Parasite Exit from Infected Erythrocytes |
| title_fullStr | Biophysics of Malarial Parasite Exit from Infected Erythrocytes |
| title_full_unstemmed | Biophysics of Malarial Parasite Exit from Infected Erythrocytes |
| title_short | Biophysics of Malarial Parasite Exit from Infected Erythrocytes |
| title_sort | biophysics of malarial parasite exit from infected erythrocytes |
| url | http://hdl.handle.net/1721.1/65418 https://orcid.org/0000-0002-6223-6831 |
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