Expression and characterization of the FXYD ion transport regulators for NMR structural studies in lipid micelles and lipid bilayers. Biochim Biophys Acta 2003 Jan 31;1645(1):15-21
Date
01/22/2003Pubmed ID
12535606Pubmed Central ID
PMC2917601DOI
10.1016/s1570-9639(02)00473-9Scopus ID
2-s2.0-0037474272 (requires institutional sign-in at Scopus site) 19 CitationsAbstract
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.
Author List
Crowell KJ, Franzin CM, Koltay A, Lee S, Lucchese AM, Snyder BC, Marassi FMAuthor
Francesca M. Marassi PhD Chair, Professor in the Biophysics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceCircular Dichroism
Cloning, Molecular
DNA, Complementary
Electrophoresis, Polyacrylamide Gel
Escherichia coli
Intracellular Signaling Peptides and Proteins
Lipid Bilayers
Lipids
Magnetic Resonance Spectroscopy
Membrane Proteins
Micelles
Molecular Sequence Data
Neoplasm Proteins
Potassium Channels
Protein Conformation
Sequence Alignment
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
