tailieunhanh - Báo cáo khoa học: Characterization of electrogenic bromosulfophthalein transport in carnation petal microsomes and its inhibition by antibodies against bilitranslocase

Bilitranslocase is a rat liver plasma membrane carrier, displaying a high-affinity binding site for bilirubin. It is competitively inhibited by grape anthocyanins, including aglycones and their mono- and di-glycosylated derivatives. In plant cells, anthocyanins are synthesized in the cytoplasm and then translocated into the central vacuole, by mechanisms yet to be fully characterized. | iFEBS Journal Characterization of electrogenic bromosulfophthalein transport in carnation petal microsomes and its inhibition by antibodies against bilitranslocase Sabina Passamonti1 Alessandra Cocolo1 Enrico Braidot2 Elisa Petrussa2 Carlo Peresson2 Nevenka Medic1 Francesco Macri2 and Angelo Vianello2 1 Dipartimento di Biochimica Biofisica e Chimica delle Macromolecole University di Trieste Italy 2 Dipartimento di Biologia ed Economia Agro-Industriale Sezione di Biologia Vegetale Universita di Udine Italy Keywords anthocyanin bilitranslocase bromosulfophthalein liver plant Correspondence S. Passamonti Dipartimento di Biochimica Biofisica e Chimica delle Macromolecole University di Trieste via L. Giorgeri 1 I-34127 Trieste Italy Fax 39 40 558 3691 Tel 39 40 558 3681 E-mail passamonti@ Website http Received 7 March 2005 revised 1 5 April 2005 accepted 5 May 2005 doi Bilitranslocase is a rat liver plasma membrane carrier displaying a high-affinity binding site for bilirubin. It is competitively inhibited by grape anthocyanins including aglycones and their mono- and di-glycosylated derivatives. In plant cells anthocyanins are synthesized in the cytoplasm and then translocated into the central vacuole by mechanisms yet to be fully characterized. The aim of this work was to determine whether a homologue of rat liver bilitranslocase is expressed in carnation petals where it might play a role in the membrane transport of anthocyanins. The bromosulfophthalein-based assay of rat liver bilitranslocase transport activity was implemented in subcellular membrane fractions leading to the identification of a bromosulfophthalein carrier KM pM which is competitively inhibited by cyanidine 3-glucoside Ki pM and mainly noncompetitively by cyanidin Ki pM . Two antisequence antibodies against bilitranslocase inhibited this carrier. In analogy to liver bilitrans-locase one antibody identified a .

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