Synthesis and spectroscopy of membrane receptor proteins. The gamma subunit of the IgE receptor.
The high-affinity receptor for IgE is a tetrameric complex of subunits of the type alpha beta gamma 2. We report here conformational studies of the intact gamma subunit in trifluoroethanol and water/liposomes by circular dichroism and Fourier-transform infrared (FTIR) spectroscopy. In trifluoroethanol, the FTIR amide I' frequencies were consistent with two predominant conformational components, the beta-turn and alpha-helix, whilst in liposomes consisting of D2O and dimyristoylglycerophosphocholine (Myr2GroPCho), three components were observed. The third component present may contain some left-handed extended helix. Spectral simulation was carried out to demonstrate that the CD spectra were consistent with the component conformations identified from FTIR spectroscopy. The stimulated CD spectra were in excellent agreement with the experimental spectra. The intact gamma subunit conformation in trifluoroethanol was shown to possess 72% alpha-helical and 28% beta-turn conformations. In water/Myr2GroPCho liposomes the percentage of each conformational component present is 37%, 38% and 25% for the alpha-helix, beta-turn and extended structures, respectively. Assuming that the transmembrane fragment was alpha-helical, an excellent correlation was found between this derived alpha-helical content in water/liposomes (37%) and from hydrophobicity plots where the percentage of amino acids in the transmembrane domain is predicted by others to be 34%. It is suggested that the beta-turn detected by CD and FTIR was attributable to a 3(10) helix rather than a type I or type III reverse turn.
Anderson
G J
GJ
University-Industry Centre for Pharmaceutical Research, University of London, England.
Haris
P I
PI
Chapman
D
D
Romer
J T
JT
Toth
G K
GK
Toth
I
I
Gibbons
W A
WA
D000944:Antigens, Differentiation, B-Lymphocyte / Q000737:chemistry*; D002942:Circular Dichroism; D005583:Fourier Analysis; D007073:Immunoglobulin E / Q000378:metabolism; D008081:Liposomes; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000138:chemical synthesis / Q000737:chemistry*; D011107:Polylysine / Q000737:chemistry; D011487:Protein Conformation; D011961:Receptors, Fc / Q000737:chemistry*; D017455:Receptors, IgE; D013055:Spectrophotometry, Infrared
D000944:Antigens, Differentiation, B-Lymphocyte; D008081:Liposomes; D046911:Macromolecular Substances; D008565:Membrane Proteins; D011961:Receptors, Fc; D017455:Receptors, IgE; D011107:Polylysine; D007073:Immunoglobulin E