Self-assembling functional programmable protein array for studying protein-protein interactions in malaria parasites

Malaria Journal
Gabriela Arévalo-PinzónManuel Fuentes

Abstract

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. 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 wel...Continue Reading

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Citations

Nov 30, 2019·Nature Reviews. Genetics·Adam D SilvermanMichael C Jewett

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Methods Mentioned

BETA
chip
Protein Arrays
PCR
protein display
Fluorescence
protein array
pull
isothermal titration calorimetry

Software Mentioned

HCIVT
AVEXIS
GenePix
AVEXYS
BLAST

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