Structure of Rhoptry Neck Protein 2 is essential for the interaction in vitro with Apical Membrane Antigen 1 in Plasmodium vivax.


Journal

Malaria journal
ISSN: 1475-2875
Titre abrégé: Malar J
Pays: England
ID NLM: 101139802

Informations de publication

Date de publication:
25 Jan 2019
Historique:
received: 01 10 2018
accepted: 13 01 2019
entrez: 27 1 2019
pubmed: 27 1 2019
medline: 8 2 2019
Statut: epublish

Résumé

In several Apicomplexa, the formation of moving junctions (MJs) at the interface between the external membranes of the invading parasite and the host cell is essential for the process of parasite invasion. In Plasmodium falciparum and Toxoplasma gondii, the MJ is composed of the Apical Membrane Antigen 1 (AMA1) and Rhoptry Neck Proteins (RONs) complex; specifically, AMA1 interacts with RON2 during host cell invasion. Recombinant proteins based on Plasmodium vivax RON2 (A2033-P2100) and its synthetic peptide fragments, one cyclic and one linear, based on PvRON2 (D2035-T2074) were generated and used to evaluate the interaction with P. vivax AMA1 (PvAMA1) by the far western blot, surface plasmon resonance (SPR), and isothermal titration microcalorimetry (ITC) methods. The structural studies of peptides were performed by circular dichroism, and the structural analysis of the complex of PvAMA1 with peptides based on PvRON2 (D2035-T2074) was conducted with small-angle X-ray scattering (SAXS). Surface plasmon resonance (KD = 23.91 ± 2.078 μmol/L) and ITC (K = 3 × 10 The results show that the PvRON2 structure, particularly the S-S bond between C2051 and C2063, is determinant for the existence of the interaction between PvAMA1 and PvRON2.

Sections du résumé

BACKGROUND BACKGROUND
In several Apicomplexa, the formation of moving junctions (MJs) at the interface between the external membranes of the invading parasite and the host cell is essential for the process of parasite invasion. In Plasmodium falciparum and Toxoplasma gondii, the MJ is composed of the Apical Membrane Antigen 1 (AMA1) and Rhoptry Neck Proteins (RONs) complex; specifically, AMA1 interacts with RON2 during host cell invasion.
METHODS METHODS
Recombinant proteins based on Plasmodium vivax RON2 (A2033-P2100) and its synthetic peptide fragments, one cyclic and one linear, based on PvRON2 (D2035-T2074) were generated and used to evaluate the interaction with P. vivax AMA1 (PvAMA1) by the far western blot, surface plasmon resonance (SPR), and isothermal titration microcalorimetry (ITC) methods. The structural studies of peptides were performed by circular dichroism, and the structural analysis of the complex of PvAMA1 with peptides based on PvRON2 (D2035-T2074) was conducted with small-angle X-ray scattering (SAXS).
RESULTS RESULTS
Surface plasmon resonance (KD = 23.91 ± 2.078 μmol/L) and ITC (K = 3 × 10
CONCLUSIONS CONCLUSIONS
The results show that the PvRON2 structure, particularly the S-S bond between C2051 and C2063, is determinant for the existence of the interaction between PvAMA1 and PvRON2.

Identifiants

pubmed: 30683104
doi: 10.1186/s12936-019-2649-6
pii: 10.1186/s12936-019-2649-6
pmc: PMC6347818
doi:

Substances chimiques

Antigens, Protozoan 0
Membrane Proteins 0
Protozoan Proteins 0
Recombinant Proteins 0
apical membrane antigen I, Plasmodium 0
rhoptry-associated antigen-2, Plasmodium 145184-78-7

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

25

Subventions

Organisme : Fundação de Amparo à Pesquisa do Estado de São Paulo
ID : 2010/17060-3
Organisme : Fundação de Amparo à Pesquisa do Estado de São Paulo
ID : 2012/13032-5
Organisme : Consejo Nacional de Innovación, Ciencia y Tecnología
ID : 21120497

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Auteurs

Perla Salgado-Mejias (P)

Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil.
Department of Chemical Sciences and Natural Resources, Faculty of Engineering and Science, University of La Frontera, Temuco, Chile.

Flavio L Alves (FL)

Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, SP, Brazil.

Kátia S Françoso (KS)

Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil.

Karin A Riske (KA)

Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, SP, Brazil.

Emerson R Silva (ER)

Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, SP, Brazil.

Antonio Miranda (A)

Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, SP, Brazil.

Irene S Soares (IS)

Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil. isoares@usp.br.

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Classifications MeSH