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

Diatloff, Eugene and Forde, Brian G. and Roberts, Stephen K. (2006) Expression and transport characterisation of the wheat low-affinity cation transporter (LCT1) in the methylotrophic yeast Pichia pastoris. Biochemical and Biophysical Research Communications, 344 (3). pp. 807-813. ISSN 0006-291X

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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.

Item Type: Article
Journal or Publication Title: Biochemical and Biophysical Research Communications
Uncontrolled Keywords: Cadmium ; Calcium ; Cation transporter ; Heterologous expression ; Low affinity ; LCT1 ; Pichia pastoris ; Triticum aestivum ; Wheat
Subjects: Q Science > QH Natural history > QH301 Biology
Departments: Faculty of Science and Technology > Lancaster Environment Centre
Faculty of Health and Medicine > Biomedical & Life Sciences
ID Code: 9510
Deposited By: Dr Stephen K Roberts
Deposited On: 12 Jun 2008 13:46
Refereed?: Yes
Published?: Published
Last Modified: 26 Jul 2012 18:38
Identification Number:
URI: http://eprints.lancs.ac.uk/id/eprint/9510

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