tetano
Editor, Senior Moderator
Protein Sci. 2012 Nov 8. doi: 10.1002/pro.2186. [Epub ahead of print]
Detection of drug-induced conformational change of a transmembrane protein in lipid bilayers using site-directed spin labeling.
Thomaston J, Nguyen PA, Brown EC, Upshur MA, Wang J, Degrado WF, Howard KP.
Source
Department of Chemistry and Biochemistry, Swarthmore College, Swarthmore, PA 19081.
Abstract
As a target of anti-viral drugs, the influenza A M2 protein has been the focus of numerous structural studies and has been extensively explored as a model ion channel. In this study we capitalize on the expanding body of 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 drug-induced conformational changes of the M2protein embedded in lipid bilayers. We obtained data in the presence of adamantane drugs for two different M2 constructs (M2TM 22-46 and M2TMC 23-60). M2TM peptides were spin-labeled at the N-terminal end of the transmembrane domain. M2TMC peptides were spin-labeled site-specifically at cysteine residues substituted for amino acids within the transmembrane domain (L36, I39, I42, L43) and the C-terminal amphipathic helix (L46, F47, F48, C50, I51, Y52, R53, F54, F55 and E56). Addition of adamantanedrugsbrought about significant changes in measured EPR environmental parameters consistent with narrowing of the transmembrane channel pore and closer packing of the C-terminal amphipathic helices.
Copyright ? 2012 The Protein Society.
PMID:
23139077
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23139077
Detection of drug-induced conformational change of a transmembrane protein in lipid bilayers using site-directed spin labeling.
Thomaston J, Nguyen PA, Brown EC, Upshur MA, Wang J, Degrado WF, Howard KP.
Source
Department of Chemistry and Biochemistry, Swarthmore College, Swarthmore, PA 19081.
Abstract
As a target of anti-viral drugs, the influenza A M2 protein has been the focus of numerous structural studies and has been extensively explored as a model ion channel. In this study we capitalize on the expanding body of 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 drug-induced conformational changes of the M2protein embedded in lipid bilayers. We obtained data in the presence of adamantane drugs for two different M2 constructs (M2TM 22-46 and M2TMC 23-60). M2TM peptides were spin-labeled at the N-terminal end of the transmembrane domain. M2TMC peptides were spin-labeled site-specifically at cysteine residues substituted for amino acids within the transmembrane domain (L36, I39, I42, L43) and the C-terminal amphipathic helix (L46, F47, F48, C50, I51, Y52, R53, F54, F55 and E56). Addition of adamantanedrugsbrought about significant changes in measured EPR environmental parameters consistent with narrowing of the transmembrane channel pore and closer packing of the C-terminal amphipathic helices.
Copyright ? 2012 The Protein Society.
PMID:
23139077
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23139077