{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,5,17]],"date-time":"2024-05-17T20:12:59Z","timestamp":1715976779656},"reference-count":70,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Microbiol"],"published-print":{"date-parts":[[2013,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>The fungus <jats:italic>Paracoccidioides spp<\/jats:italic> is the agent of paracoccidioidomycosis (PCM), a pulmonary mycosis acquired by the inhalation of fungal propagules. <jats:italic>Paracoccidioides<\/jats:italic> malate synthase (<jats:italic>Pb<\/jats:italic>MLS) is important in the infectious process of <jats:italic>Paracoccidioides spp<\/jats:italic> because the transcript is up-regulated during the transition from mycelium to yeast and in yeast cells during phagocytosis by murine macrophages. In addition, <jats:italic>Pb<\/jats:italic>MLS acts as an adhesin in <jats:italic>Paracoccidioides spp<\/jats:italic>. The evidence for the multifunctionality of <jats:italic>Pb<\/jats:italic>MLS indicates that it could interact with other proteins from the fungus and host. The objective of this study was to identify and analyze proteins that possibly bind to <jats:italic>Pb<\/jats:italic>MLS (<jats:italic>Pb<\/jats:italic>MLS-interacting proteins) because protein interactions are intrinsic to cell processes, and it might be possible to infer the function of a protein through the identification of its ligands.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>The search for interactions was performed using an <jats:italic>in vivo<\/jats:italic> assay with a two-hybrid library constructed in <jats:italic>S. cerevisiae<\/jats:italic>; the transcripts were sequenced and identified. In addition, an <jats:italic>in vitro<\/jats:italic> assay using pull-down GST methodology with different protein extracts (yeast, mycelium, yeast-secreted proteins and macrophage) was performed, and the resulting interactions were identified by mass spectrometry (MS). Some of the protein interactions were confirmed by Far-Western blotting using specific antibodies, and the interaction of <jats:italic>Pb<\/jats:italic>MLS with macrophages was validated by indirect immunofluorescence and confocal microscopy. <jats:italic>In silico<\/jats:italic> analysis using molecular modeling, dynamics and docking identified the amino acids that were involved in the interactions between <jats:italic>Pb<\/jats:italic>MLS and <jats:italic>Pb<\/jats:italic>MLS-interacting proteins. Finally, the interactions were visualized graphically using Osprey software.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>These observations indicate that <jats:italic>Pb<\/jats:italic>MLS interacts with proteins that are in different functional categories, such as cellular transport, protein biosynthesis, modification and degradation of proteins and signal transduction. These data suggest that <jats:italic>Pb<\/jats:italic>MLS could play different roles in the fungal cell.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1471-2180-13-107","type":"journal-article","created":{"date-parts":[[2013,5,14]],"date-time":"2013-05-14T18:16:01Z","timestamp":1368555361000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Intermolecular interactions of the malate synthase of Paracoccidioides spp"],"prefix":"10.1186","volume":"13","author":[{"given":"Karine Martins","family":"de Oliveira","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benedito Rodrigues","family":"da Silva Neto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juliana Alves","family":"Parente","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roosevelt Alves","family":"da Silva","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guilherme Oliveira","family":"Quintino","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aline Raquel","family":"Voltan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Jos\u00e9 Soares","family":"Mendes-Giannini","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C\u00e9lia Maria","family":"de Almeida Soares","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maristela","family":"Pereira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2013,5,14]]},"reference":[{"key":"1986_CR1","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1128\/CMR.6.2.89","volume":"6","author":"E Brummer","year":"1993","unstructured":"Brummer E, Castaneda E, Restrepo A: Paracoccidioidomycosis: an update. 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