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A substrate translocation trajectory in a cytoplasm-facing topological model of...
Silva, Isabel Soares; Pessoa, Joana Sá; Myrianthopoulos, Vassilios; Mikros, Emmanuel; Casal, Margarida; Diallinas, GeorgePrevious mutational analysis of Jen1p, a Saccharomyces cerevisiae monocarboxylate/H+ symporter of the Major Facilitator Superfamily, has suggested that the consensus sequence 379NXX[S/T]HX[S/T]QD387 in transmembrane segment VII (TMS-VII) is part of the substrate translocation pathway. Here, we rationally design, analyse and show that several novel mutations in TMS-V and TMS-XI directly modify Jen1p function. Am...
A substrate translocation trajectory in the monocarboxylate/h+ symporter jen1
Pessoa, Joana Sá; Silva, Isabel Soares; Myrianthopoulos, Vassilios; Mikros, Emmanuel; Diallinas, George; Casal, MargaridaPrevious mutational analysis of Jen1p, a Saccharomyces cerevisiae monocarboxylate/H+ symporter of the Major Facilitator Superfamily, has suggested that the consensus sequence 379NXX[S/T]HX[S/T]QD387, located in transmembrane segment VII (TMS-VII), is part of the substrate translocation pathway. In this work, we rationally design and analyse novel mutations concerning residues in TMS-V and TMS-XI. Our analysis i...
Identification of amino acid residues critical for the substrate translocation ...
Pessoa, Joana Sá; Silva, Isabel Soares; Myrianthopoulos, Vassilios; Mikros, Emmanuel; Diallinas, George; Casal, MargaridaLactic, acetic and propionic acids have been used for many years in industrial and pharmaceutical companies. In Saccharomyces cerevisiae, Jen1p is a major monocarboxylate:H+ symporter specific primarily for lactate, pyruvate and for acetate (TC # 2.A.1.12.2) (Casal et al., 1999). A phylogenetic tree of ScJen1p homologues (Casal et al., 2008) showed the existence of two main clusters: a Jen1 group (monocarboxyl...
Identification of amino acid residues critical for distinguishing mono- and di-...
Pessoa, Joana Sá; Silva, Isabel Soares; Diallinas, George; Casal, MargaridaLactic, acetic and propionic acids have been used for many years in industrial and pharmaceutical companies and, more recently, lactate as been used for production of biodegradable polymers and as substitute for petroleum-derived chemicals. Understanding in detail the mechanisms underlying the transport of carboxylic acids is crucial towards an efficient biological production of these compounds. In Saccharomyce...
Identification of amino acid residues critical for distinguishing mono- and di-...
Pessoa, Joana Sá; Silva, Isabel Soares; Diallinas, George; Casal, MargaridaThe knowledge of the mechanisms underlying the transport of carboxylic acids is crucial towards an efficient biological production of carboxylates which have been used for many years in industry namely for the production of biodegradable polymers and as substitute for petroleum-derived chemicals. In <i>Saccharomyces cerevisiae</i>, Jen1p is major monocarboxylate H+ symporter specific primarily for lactate, pyru...
Characterization of monocarboxylate permeases in yeast : functional and structu...
Silva, Isabel SoaresOs ácidos carboxílicos de cadeia curta são compostos importantes produzidos de forma natural no decorrer de numerosos processos metabólicos, para além de poderem ser utilizados como única fonte de carbono e energia por diversos seres vivos, incluindo leveduras. É neste contexto que o estudo das permeases de monocarboxilatos se reveste de especial importância, dado que o transporte destes nutrientes através da m...
The disruption of JEN1 from Candida albicans impairs the transport of lactate
Paiva, Sandra; Silva, Isabel Soares; Kötter, Peter; Entian, Karl-Dieter; Casal, MargaridaPoster apresentado em XXI International Conference on Yeast Genetics and Molecular Biology, Gotemburgo, Suécia, 7 Jul. - 12 Jul. 2004
The disruption of JEN1 from Candida albicans impairs the transport of lactate
Silva, Isabel Soares; Paiva, Sandra; Kötter, Peter; Entian, Karl-Dieter; Casal, MargaridaA lactate permease was biochemically identified in Candida albicans RM1000 presenting the following kinetic parameters at pH 5.0: Km 0.33 ± 0.09 mM and Vmax 0.85± 0.06 nmol s-1 mg dry wt-1. Lactate uptake was competitively inhibited by pyruvic and propionic acids; acetic acid behaved as a non-competitive substrate. An ORF homologous to Saccharomyces cerevisiae gene JEN1 was identified (CaJEN1). Deletions of bot...
Functional expression of the lactate permease Jen1p of Saccharomyces cerevisiae...
Silva, Isabel Soares; Schuller, Dorit; Andrade, Raquel P.; Baltazar, Fátima; Cássio, Fernanda; Casal, MargaridaIn Saccharomyces cerevisiae the activity for the lactate proton-symporter in dependent on JEN1 gene expression. Pichia pastoris was transformed with an integrative plasmid containing the JEN1 gene. After 24 h of methanol induction, Northern and Western-blotting analyses indicated the expression of JEN1 in the transformants. Lactate permease activity was obtained in P. pastoris cells with a Vmax of 2.1 nmol s-1 ...
Exploring genetic tools for the overexpression of the lactate permease Jen1p of...
Silva, Isabel Soares; Andrade, Raquel P.; Schuller, Dorit; Casal, MargaridaResumo e poster da comunicação apresentada sob forma de painel no congresso "XXI International Conference on Yeast Genetics and Molecular Biology", em 2003, em Göteborg, Suécia. ; In Saccharomyces cerevisiae the active transport of monocarboxylic acids is dependent upon the expression of the permease Jen1p. Cells of S. cerevisiae jen1 were transformed with the plasmids p416GPD (centromeric) and p426GPD (multic...
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