Structural flexibility of the pentameric SARS coronavirus envelope protein ion channel.
Coronaviruses contain a small envelope membrane protein with cation-selective ion channel activity mediated by its transmembrane domain (ETM). In a computational study, we proposed that ion channel activity can be explained by either of two similar ETM homopentameric transmembrane alpha-helical bundles, related by a approximately 50 degrees rotation of the helices. Later, we tested this prediction, using site-specific infrared dichroism of a lysine-flanked isotopically labeled ETM peptide from the virus responsible for the severe acute respiratory syndrome, SARS, reconstituted in lipid bilayers. However, the data were consistent with the presence of a kink at the center of the ETM alpha-helix, and it did not fit completely either computational model. Herein, we have used native ETM, without flanking lysines, and show that the helix orientation is now consistent with one of the predicted models. ETM only produced one oligomeric form, pentamers, in the lipid-mimic detergent dodecylphosphocholine and in perfluorooctanoic acid. We thus report the correct backbone model for the pentameric alpha-helical bundle of ETM. The disruptive effects caused by terminal lysines probably highlight the conformational flexibility required during ion channel function.
Parthasarathy
Krupakar
K
Ng
Lifang
L
Lin
Xin
X
Liu
Ding Xiang
DX
Pervushin
Konstantin
K
Gong
Xiandi
X
Torres
Jaume
J
D000595:Amino Acid Sequence; D000818:Animals; D002210:Caprylates / Q000737:chemistry; D004586:Electrophoresis; D005466:Fluorocarbons / Q000737:chemistry; D006801:Humans; D007473:Ion Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D045473:Severe acute respiratory syndrome-related coronavirus / Q000737:chemistry*; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D002210:Caprylates; D005466:Fluorocarbons; D007473:Ion Channels; D014759:Viral Envelope Proteins; C023036:perfluorooctanoic acid