NDLI: Characterization of a calmodulin-binding transporter from the plasma membrane of barley aleurone

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Characterization of a calmodulin-binding transporter from the plasma membrane of barley aleurone

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Characterization of a calmodulin-binding transporter from the plasma membrane of barley aleurone

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Shartzer, S. F. Schuurink, R. C. Jones, R. L. Fath, A.
Description	Author Affiliation: Schuurink RC ( Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720-3102, USA.);
Abstract	We have used Arabidopsis calmodulin (CaM) covalently coupled to horseradish peroxidase to screen a barley aleurone cDNA expression library for CaM binding proteins. The deduced amino acid sequence of one cDNA obtained by this screen was shown to be a unique protein of 702 amino acids with CaM and cyclic nucleotide binding domains at the carboxyl terminus and high similarity to olfactory and K+ channels. This cDNA was designated HvCBT1 (Hordeum vulgare CaM binding transporter). Hydropathy plots of HvCBT1 showed the presence of six putative transmembrane domains, but sequence alignment indicated a pore domain that was unlike the consensus domains in K+ and olfactory channels. Expression of a subclone of amino acids 482-702 in Escherichia coli generated a peptide that bound CaM. When a fusion protein of HvCBT1 and green fluorescent protein was expressed in barley aleurone protoplasts, fluorescence accumulated in the plasma membrane. Expression of HvCBT1 in the K+ transport deficient Saccharomyces cerevisiae mutant CY162 showed no rescue of the mutant phenotype. However, growth of CY162 expressing HvCBT1 with its pore mutated to GYGD, the consensus sequence of K+ channels, was compromised. We interpret these data as indicating that HvCBT1 acts to interfere with ion transport.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	4
Volume Number	95
Language	English
Publisher	National Academy of Sciences
Publisher Date	1998-03-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Calmodulin-Binding Proteins Isolation & Purification Hordeum Chemistry Membrane Proteins Plant Proteins Amino Acid Sequence Biological Transport Genetics Cell Membrane Cloning, Molecular DNA, Complementary Ion Channels Molecular Sequence Data Research Support, U.S. Gov't, Non-P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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