title: Determining the Secondary Structure of Membrane Proteins and Peptides Via Electron Spin Echo Envelope Modulation (ESEEM) Spectroscopy.
abstract: Revealing detailed structural and dynamic information of membrane embedded or associated proteins is challenging due to their hydrophobic nature which makes NMR and X-ray crystallographic studies challenging or impossible. Electron paramagnetic resonance (EPR) has emerged as a powerful technique to provide essential structural and dynamic information for membrane proteins with no size limitations in membrane systems which mimic their natural lipid bilayer environment. Therefore, tremendous efforts have been devoted toward the development and application of EPR spectroscopic techniques to study the structure of biological systems such as membrane proteins and peptides. This chapter introduces a novel approach established and developed in the Lorigan lab to investigate membrane protein and peptide local secondary structures utilizing the pulsed EPR technique electron spin echo envelope modulation (ESEEM) spectroscopy. Detailed sample preparation strategies in model membrane protein systems and the experimental setup are described. Also, the ability of this approach to identify local secondary structure of membrane proteins and peptides with unprecedented efficiency is demonstrated in model systems. Finally, applications and further developments of this ESEEM approach for probing larger size membrane proteins produced by overexpression systems are discussed.
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, USA. Electronic address: lorigag@miamioh.edu.
authors.forename: Lishan
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, USA.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Mayo
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, USA.
authors.forename: Indra D
authors.initials: ID
authors.lastname: Sahu
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, USA.
authors.forename: Andy
authors.initials: A
authors.lastname: Zhou
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, USA.
authors.forename: Rongfu
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: McCarrick
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, USA.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Lorigan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D017433:Protein Structure, Secondary; D013113:Spin Labels
chemical_list: D008565:Membrane Proteins; D010455:Peptides; D013113:Spin Labels
keywords: ESEEM; Membrane peptide; Membrane protein; Pulsed EPR; SDSL; Secondary structure