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By the first approach, Coomassie-stained protein bands (32- and 33-kDa fractions) that specifically reacted by immunoblotting with sera from MVL patients were excised from the gel and submitted to enzymatic digestion to generate peptides. Four peptides were sequenced, three of which were shown to be definitely associated with MVL-reactive antigens and ascribed to a mitochondrial integral ADP-ATP carrier protein from<jats:italic>L. major<\/jats:italic>, a putative NADH cytochrome<jats:italic>b<\/jats:italic><jats:sub>5<\/jats:sub>reductase, and a putative mitochondrial carrier protein, respectively. The second approach combined two-dimensional gel electrophoresis of membrane antigens and mass spectrometry (liquid chromatography-mass spectrometry\/mass spectrometry) by using a quadrupole time-of-flight analysis. Six immunoreactive spots that resolved within a molecular mass range of 30 to 36 kDa and a pH range of 6.7 to 7.4 corresponded to four<jats:italic>Leishmania<\/jats:italic>products. The sequences derived from two spots were ascribed to a beta subunit-like guanine nucleotide binding protein, known as the activated protein kinase C receptor homolog antigen LACK, and to a probable member of the aldehyde reductase family. One spot was identified as a probable ubiquinol-cytochrome<jats:italic>c<\/jats:italic>reductase (EC 1.10.2.2) Rieske iron-sulfur protein precursor. The remaining three spots were identified as truncated forms of elongation factor 1\u03b1. These antigens correspond to conserved proteins ubiquitously expressed in eukaryotic cells and represent potential candidates for the design of a reliable tool for the diagnosis of this disease.<\/jats:p>","DOI":"10.1128\/cdli.12.2.310-320.2005","type":"journal-article","created":{"date-parts":[[2005,2,7]],"date-time":"2005-02-07T22:28:47Z","timestamp":1107815327000},"page":"310-320","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":10,"title":["Proteomic Approach for Characterization of Immunodominant Membrane-Associated 30- to 36-Kilodalton Fraction Antigens of<i>Leishmania infantum<\/i>Promastigotes, Reacting with Sera from Mediterranean Visceral Leishmaniasis Patients"],"prefix":"10.1128","volume":"12","author":[{"given":"Sayda","family":"Kamoun-Essghaier","sequence":"first","affiliation":[{"name":"Laboratoire d'Immunologie, Vaccinologie et Ge\u0301ne\u0301tique Mole\u0301culaire"},{"name":"Laboratoire 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