Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites
Abstract Background Plasmodium vivax is the most widespread malarial species, causing significant morbidity worldwide. Knowledge is limited regarding the molecular mechanism of invasion due to the lack of a continuous in vitro culture system for these species. Since protein–protein and host–cell int...
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ftdoajarticles:oai:doaj.org/article:aa03c72603bf43d880362d5fad0bd8ba 2023-05-15T15:10:29+02:00 Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites Gabriela Arévalo-Pinzón María González-González Carlos Fernando Suárez Hernando Curtidor Javier Carabias-Sánchez Antonio Muro Joshua LaBaer Manuel Alfonso Patarroyo Manuel Fuentes 2018-07-01T00:00:00Z https://doi.org/10.1186/s12936-018-2414-2 https://doaj.org/article/aa03c72603bf43d880362d5fad0bd8ba EN eng BMC http://link.springer.com/article/10.1186/s12936-018-2414-2 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-018-2414-2 1475-2875 https://doaj.org/article/aa03c72603bf43d880362d5fad0bd8ba Malaria Journal, Vol 17, Iss 1, Pp 1-14 (2018) Plasmodium vivax Malaria NAPPA array IVTT protein expression Protein–protein interaction Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2018 ftdoajarticles https://doi.org/10.1186/s12936-018-2414-2 2022-12-31T01:33:13Z Abstract Background Plasmodium vivax is the most widespread malarial species, causing significant morbidity worldwide. Knowledge is limited regarding the molecular mechanism of invasion due to the lack of a continuous in vitro culture system for these species. Since protein–protein and host–cell interactions play an essential role in the microorganism’s invasion and replication, elucidating protein function during invasion is critical when developing more effective control methods. Nucleic acid programmable protein array (NAPPA) has thus become a suitable technology for studying protein–protein and host–protein interactions since producing proteins through the in vitro transcription/translation (IVTT) method overcomes most of the drawbacks encountered to date, such as heterologous protein production, stability and purification. Results Twenty P. vivax proteins on merozoite surface or in secretory organelles were selected and successfully cloned using gateway technology. Most constructs were displayed in the array expressed in situ, using the IVTT method. The Pv12 protein was used as bait for evaluating array functionality and co-expressed with P. vivax cDNA display in the array. It was found that Pv12 interacted with Pv41 (as previously described), as well as PvMSP142kDa, PvRBP1a, PvMSP8 and PvRAP1. Conclusions NAPPA is a high-performance technique enabling co-expression of bait and query in situ, thereby enabling interactions to be analysed rapidly and reproducibly. It offers a fresh alternative for studying protein–protein and ligand–receptor interactions regarding a parasite which is difficult to cultivate (i.e. P. vivax). Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 17 1 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Plasmodium vivax Malaria NAPPA array IVTT protein expression Protein–protein interaction Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Plasmodium vivax Malaria NAPPA array IVTT protein expression Protein–protein interaction Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Gabriela Arévalo-Pinzón María González-González Carlos Fernando Suárez Hernando Curtidor Javier Carabias-Sánchez Antonio Muro Joshua LaBaer Manuel Alfonso Patarroyo Manuel Fuentes Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites |
topic_facet |
Plasmodium vivax Malaria NAPPA array IVTT protein expression Protein–protein interaction Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Plasmodium vivax is the most widespread malarial species, causing significant morbidity worldwide. Knowledge is limited regarding the molecular mechanism of invasion due to the lack of a continuous in vitro culture system for these species. Since protein–protein and host–cell interactions play an essential role in the microorganism’s invasion and replication, elucidating protein function during invasion is critical when developing more effective control methods. Nucleic acid programmable protein array (NAPPA) has thus become a suitable technology for studying protein–protein and host–protein interactions since producing proteins through the in vitro transcription/translation (IVTT) method overcomes most of the drawbacks encountered to date, such as heterologous protein production, stability and purification. Results Twenty P. vivax proteins on merozoite surface or in secretory organelles were selected and successfully cloned using gateway technology. Most constructs were displayed in the array expressed in situ, using the IVTT method. The Pv12 protein was used as bait for evaluating array functionality and co-expressed with P. vivax cDNA display in the array. It was found that Pv12 interacted with Pv41 (as previously described), as well as PvMSP142kDa, PvRBP1a, PvMSP8 and PvRAP1. Conclusions NAPPA is a high-performance technique enabling co-expression of bait and query in situ, thereby enabling interactions to be analysed rapidly and reproducibly. It offers a fresh alternative for studying protein–protein and ligand–receptor interactions regarding a parasite which is difficult to cultivate (i.e. P. vivax). |
format |
Article in Journal/Newspaper |
author |
Gabriela Arévalo-Pinzón María González-González Carlos Fernando Suárez Hernando Curtidor Javier Carabias-Sánchez Antonio Muro Joshua LaBaer Manuel Alfonso Patarroyo Manuel Fuentes |
author_facet |
Gabriela Arévalo-Pinzón María González-González Carlos Fernando Suárez Hernando Curtidor Javier Carabias-Sánchez Antonio Muro Joshua LaBaer Manuel Alfonso Patarroyo Manuel Fuentes |
author_sort |
Gabriela Arévalo-Pinzón |
title |
Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites |
title_short |
Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites |
title_full |
Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites |
title_fullStr |
Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites |
title_full_unstemmed |
Self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites |
title_sort |
self-assembling functional programmable protein array for studying protein–protein interactions in malaria parasites |
publisher |
BMC |
publishDate |
2018 |
url |
https://doi.org/10.1186/s12936-018-2414-2 https://doaj.org/article/aa03c72603bf43d880362d5fad0bd8ba |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 17, Iss 1, Pp 1-14 (2018) |
op_relation |
http://link.springer.com/article/10.1186/s12936-018-2414-2 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-018-2414-2 1475-2875 https://doaj.org/article/aa03c72603bf43d880362d5fad0bd8ba |
op_doi |
https://doi.org/10.1186/s12936-018-2414-2 |
container_title |
Malaria Journal |
container_volume |
17 |
container_issue |
1 |
_version_ |
1766341514851516416 |