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The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function
Plasmepsin V (PM V) is a pepsin-like aspartic protease essential for growth of the malarial parasite Plasmodium falciparum. Previous work has shown PM V to be an endoplasmic reticulum–resident protease that processes parasite proteins destined for export into the host cell. Depletion or inhibition o...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9478907/ https://www.ncbi.nlm.nih.gov/pubmed/35952758 http://dx.doi.org/10.1016/j.jbc.2022.102355 |
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author | Polino, Alexander J. Miller, Justin J. Bhakat, Soumendranath Mukherjee, Sumit Bobba, Suhas Bowman, Gregory R. Goldberg, Daniel E. |
author_facet | Polino, Alexander J. Miller, Justin J. Bhakat, Soumendranath Mukherjee, Sumit Bobba, Suhas Bowman, Gregory R. Goldberg, Daniel E. |
author_sort | Polino, Alexander J. |
collection | PubMed |
description | Plasmepsin V (PM V) is a pepsin-like aspartic protease essential for growth of the malarial parasite Plasmodium falciparum. Previous work has shown PM V to be an endoplasmic reticulum–resident protease that processes parasite proteins destined for export into the host cell. Depletion or inhibition of the enzyme is lethal during asexual replication within red blood cells as well as during the formation of sexual stage gametocytes. The structure of the Plasmodium vivax PM V has been characterized by X-ray crystallography, revealing a canonical pepsin fold punctuated by structural features uncommon to secretory aspartic proteases; however, the function of this unique structure is unclear. Here, we used parasite genetics to probe these structural features by attempting to rescue lethal PM V depletion with various mutant enzymes. We found an unusual nepenthesin 1-type insert in the PM V gene to be essential for parasite growth and PM V activity. Mutagenesis of the nepenthesin insert suggests that both its amino acid sequence and one of the two disulfide bonds that undergird its structure are required for the insert’s role in PM V function. Furthermore, molecular dynamics simulations paired with Markov state modeling suggest that mutations to the nepenthesin insert may allosterically affect PM V catalysis through multiple mechanisms. Taken together, these data provide further insights into the structure of the P. falciparum PM V protease. |
format | Online Article Text |
id | pubmed-9478907 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-94789072022-09-22 The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function Polino, Alexander J. Miller, Justin J. Bhakat, Soumendranath Mukherjee, Sumit Bobba, Suhas Bowman, Gregory R. Goldberg, Daniel E. J Biol Chem Research Article Plasmepsin V (PM V) is a pepsin-like aspartic protease essential for growth of the malarial parasite Plasmodium falciparum. Previous work has shown PM V to be an endoplasmic reticulum–resident protease that processes parasite proteins destined for export into the host cell. Depletion or inhibition of the enzyme is lethal during asexual replication within red blood cells as well as during the formation of sexual stage gametocytes. The structure of the Plasmodium vivax PM V has been characterized by X-ray crystallography, revealing a canonical pepsin fold punctuated by structural features uncommon to secretory aspartic proteases; however, the function of this unique structure is unclear. Here, we used parasite genetics to probe these structural features by attempting to rescue lethal PM V depletion with various mutant enzymes. We found an unusual nepenthesin 1-type insert in the PM V gene to be essential for parasite growth and PM V activity. Mutagenesis of the nepenthesin insert suggests that both its amino acid sequence and one of the two disulfide bonds that undergird its structure are required for the insert’s role in PM V function. Furthermore, molecular dynamics simulations paired with Markov state modeling suggest that mutations to the nepenthesin insert may allosterically affect PM V catalysis through multiple mechanisms. Taken together, these data provide further insights into the structure of the P. falciparum PM V protease. American Society for Biochemistry and Molecular Biology 2022-08-09 /pmc/articles/PMC9478907/ /pubmed/35952758 http://dx.doi.org/10.1016/j.jbc.2022.102355 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Polino, Alexander J. Miller, Justin J. Bhakat, Soumendranath Mukherjee, Sumit Bobba, Suhas Bowman, Gregory R. Goldberg, Daniel E. The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function |
title | The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function |
title_full | The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function |
title_fullStr | The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function |
title_full_unstemmed | The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function |
title_short | The nepenthesin insert in the Plasmodium falciparum aspartic protease plasmepsin V is necessary for enzyme function |
title_sort | nepenthesin insert in the plasmodium falciparum aspartic protease plasmepsin v is necessary for enzyme function |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9478907/ https://www.ncbi.nlm.nih.gov/pubmed/35952758 http://dx.doi.org/10.1016/j.jbc.2022.102355 |
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