Home General Physiology and Biophysics 2013 General Physiology and Biophysics Vol.32, No.4, p.479-488, 2013

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Quarterly, 80 pp. per issue
Founded: 1982
ISSN  1338-4325 (online)

Published in English

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General Physiology and Biophysics Vol.32, No.4, p.479-488, 2013

Title: Interactions of y+LAT1 and 4F2hc in the y+l amino acid transporter complex: consequences of lysinuric protein intolerance-causing mutations
Author: Minna Toivonen,, Maaria Tringham, Johanna Kurko, Perttu Terho, Olli Simell, Kaisa M. Heiskanen, Juha Mykkänen

Abstract: Lysinuric protein intolerance (LPI) is an inherited aminoaciduria caused by recessive mutations in the SLC7A7 gene encoding y+L amino acid transporter 1 (y+LAT1), which combines with 4F2hc to generate an active transporter responsible for the system y+L amino acid transport. We have previously shown that the y+LAT1 proteins with point mutations are expressed in the plasma membrane, while those with frame shift mutations are retained in the cytoplasm. This finding has prompted us to study whether the difference in localization is due to the inability of the structurally altered mutant y+LAT1 proteins to heteromerize with 4F2hc. For this purpose, we utilized FACS technique to reveal fluorescence resonance energy transfer (FRET) in cells expressing wild type or LPI-mutant CFP-tagged y+LAT1 and YFP-tagged 4F2hc. The heteromerization of y+LAT1 and 4F2hc within the cell is not disrupted by any of the tested LPI mutations. In addition, the expression rate of the LPI mutant y+LAT1 proteins was significantly lower and cellular mortality was markedly increased than that of the wild type y+LAT1 in transfected samples. Our results indicate that the FACS-FRET method provides an alternative approach for screening of potential protein associations.

Keywords: Fluorescence activated cell sorting (FACS) — Fluorescence resonance energy transfer (FRET) — Heteromerization — Lysinuric protein intolerance (LPI) — y+LAT1
Published online: 26-Aug-2013
Year: 2013, Volume: 32, Issue: 4 Page From: 479, Page To: 488

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