Expression and characterization of the FXYD ion transport regulators for NMR structural studies in lipid micelles and lipid bilayers.

Abstract	The proteins PLM (phospholemman), CHIF (channel inducing factor), and Mat8 (mammary tumor protein 8 kDa) are members of the FXYD family of ion transport regulatory membrane proteins. Here we describe their cloning and expression in Escherichia coli, and their purification for NMR structural studies in lipid micelles and lipid bilayers. The molecular masses of the purified recombinant FXYD proteins, determined from SDS-PAGE and from MALDI TOF mass spectrometry, reflect monomeric species. The solution NMR and CD spectra in SDS micelles show that they adopt helical conformations. The solid-state NMR spectra in lipid bilayers give the first view of their transmembrane architecture.
Authors	Kevin J Crowell, Carla M Franzin, Anita Koltay, Sangmin Lee, Anna Maria Lucchese, Bradley C Snyder, Francesca M Marassi
Journal	Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 1645 Issue 1 Pg. 15-21 (Jan 31 2003) ISSN: 0006-3002 [Print] Netherlands
PMID	12535606 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)
Chemical References	DNA, Complementary FXYD3 protein, human Fxyd4 protein, rat Intracellular Signaling Peptides and Proteins Lipid Bilayers Lipids Membrane Proteins Micelles Neoplasm Proteins Potassium Channels
Topics	Amino Acid Sequence Circular Dichroism Cloning, Molecular DNA, Complementary (biosynthesis, chemistry) Electrophoresis, Polyacrylamide Gel Escherichia coli (metabolism) Intracellular Signaling Peptides and Proteins Lipid Bilayers (chemistry) Lipids (chemistry) Magnetic Resonance Spectroscopy Membrane Proteins (biosynthesis, chemistry, genetics) Micelles Molecular Sequence Data Neoplasm Proteins (biosynthesis, chemistry, genetics) Potassium Channels (biosynthesis, chemistry, genetics) Protein Conformation Sequence Alignment Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

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