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J Proteome Res ; 18(11): 3831-3839, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31549843

RESUMO

Plasmodium falciparum variant antigens named erythrocyte membrane protein 1 (PfEMP1) are important targets for developing a protective immunity to malaria caused by P. falciparum. One of the major challenges in P. falciparum proteomics studies is identifying PfEMP1s at the protein level due to antigenic variation. To identify these PfEMP1s using shotgun proteomics, we developed a pipeline that searches high-resolution mass spectrometry spectra against a custom protein sequence database. A local alignment algorithm, LAX, was developed as a part of the pipeline that matches peptide sequences to the most similar PfEMP1 and calculates a weight value based on peptide's uniqueness used for PfEMP1 protein inference. The pipeline was first validated in the analysis of a laboratory strain with a known PfEMP1, then it was implemented on the analysis of parasite isolates from malaria-infected pregnant women and finally on the analysis of parasite isolates from malaria-infected children where there was an increase of PfEMP1s identified in 27 out of 31 isolates using the expanded database.


Assuntos
Proteínas Mutantes/metabolismo , Plasmodium falciparum/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Proteínas de Protozoários/metabolismo , Sequência de Aminoácidos , Criança , Cromatografia Líquida/métodos , Feminino , Humanos , Malária Falciparum/parasitologia , Proteínas Mutantes/genética , Plasmodium falciparum/isolamento & purificação , Plasmodium falciparum/fisiologia , Gravidez , Complicações Parasitárias na Gravidez/parasitologia , Proteoma/genética , Proteínas de Protozoários/genética , Homologia de Sequência de Aminoácidos , Espectrometria de Massas em Tandem/métodos
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