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Expression and transport characterisation of the wheat low-affinity cation transporter (LCT1) in the methylotrophic yeast Pichia pastoris.

Research output: Contribution to journalJournal article

Published

Journal publication date9/06/2006
JournalBiochemical and Biophysical Research Communications
Journal number3
Volume344
Number of pages7
Pages807-813
Original languageEnglish

Abstract

The low-affinity cation transporter (LCT1) from wheat (Triticum aestivum) was expressed in the methylotrophic yeast Pichia pastoris and its transport characteristics studied employing Ca45 and Cd109. A clone (LCT1#3) with the highest uptake of 14 pmol of Ca/106 cells/10 min when exposed to 100 μM Ca45 was chosen for further Ca45 and Cd109 transport characteristics. We report for the first time a Km for Ca by LCT1 of 0.43 ± 0.15 mM Ca activity which confirms LCT1 to be a low affinity transporter. Interestingly, the expression of LCT1 in Pichia resulted in reduced Cd109 uptake compared to wild type cells, when cells were exposed to 60 μM Cd. This is the first report of the ability of a heterologously expressed transporter to reduce the activity of endogenous transporter proteins to transport Cd. To our knowledge, this is the first demonstration of functional expression of a plant ion transporter using P. pastoris.