TAILIEUCHUNG - Báo cáo khoa học: Uptake of bilirubin into HepG2 cells assayed by thermal lens spectroscopy Function of bilitranslocase

Bilitranslocase is a carrier protein localized at the basolateral domain of the hepatocyte plasma membrane. It transports various organic anions, including bromosulfophthalein and anthocyanins. Functional studies in subcellular fractions enriched in plasma membrane revealed a high-affinity binding site for bilirubin, associated with bilitranslocase. | iFEBS Journal Uptake of bilirubin into HepG2 cells assayed by thermal lens spectroscopy Function of bilitranslocase Sabina Passamonti1 Michela Terdoslavich1 Alja Margon1 2 Alessandra Cocolo1 Nevenka Medic1 Fulvio Micali1 Giuliana Decorti3 and Mladen Franko2 1 Dipartimento di Biochimica Biofisica e Chimica delle Macromolecole Universita di Trieste Italy 2 Laboratory for EnvironmentalResearch Nova Gorica Polytechnic Slovenia 3 Dipartimento di Scienze Biomediche Universita di Trieste Italy Keywords bilirubin bilitranslocase HepG2 cells thermal lens spectrometry uptake assay Correspondence S. Passamonti Dipartimento di Biochimica Biofisica e Chimica delle Macromolecole University di Trieste via L. Giorgeri 1 I-34127 Trieste Italy Fax 39 040 558 3691 Tel 39 040 558 3681 E-mail passamonti@ URL http Received 28 July 2005 revised 18 August 2005 accepted 30 August 2005 doi Bilitranslocase is a carrier protein localized at the basolateral domain of the hepatocyte plasma membrane. It transports various organic anions including bromosulfophthalein and anthocyanins. Functional studies in subcellular fractions enriched in plasma membrane revealed a high-affinity binding site for bilirubin associated with bilitranslocase. The aim of this work was to test whether the liver uptake of bilirubin depends on the activity of bilitranslocase. To this purpose an assay of bilirubin uptake into HepG2 cell cultures was set up. The transport assay medium contained bilirubin at a concentration of w 50 nM in the absence of albumin. To analyse the relative changes in bilirubin concentration in the medium throughout the uptake experiment a highly sensitive thermal lens spectrometry method was used. The mechanism of bilirubin uptake into HepG2 cells was investigated by using inhibitors such as anti-sequence bilitranslocase antibodies the protein-modifying reagent phenylmethanesulfonyl fluoride and diverse organic anions including .

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