Site Directed Spin Label EPR Spectroscopy Of Influenza A M2 Protein
M2 is a membrane protein critical to the life cycle of influenza A. We have capitalized on the expanding body of biochemical and high-resolution structural data available for the M2 protein to design and interpret site-directed spin labeling electron paramagnetic resonance spectroscopy (SDSL-EPR) experiments on the conformation and dynamics of the homotetrameric M2 protein embedded in lipid bilayers. We have obtained data for three different M2 constructs (M2TM 22-46, M2TMC 23-60 and full length M2 protein) spin-labeled at multiple sites within the transmembrane and C-terminal domains. CW and pulsed EPR spectra show evidence that M2 adopts multiple conformational states in bilayers, and that cholesterol content dictates the relative populations of the states. Furthermore comparison of full-length M2 protein and a M2TMC 23-60 peptide demonstrates that the C-terminal juxtamembrane region in both constructs forms an amphipathic membrane surface helix.
ACERT Workshop 2015: "Computational Methods In ESR"
June 9-12, 2015
Kathleen P. Howard; Bryan M. Green , '16; Shenstone Chia Huang , '16; and Sand Woo Kim , '14.
"Site Directed Spin Label EPR Spectroscopy Of Influenza A M2 Protein".
Issue 2, Suppl. 1.