Preparation of insoluble transmembrane peptides: Glycophorin-A, prion (110-137), and FGFR (368-397)

A general procedure for the reliable preparation of large quantities of ins oluble transmembrane peptides has been developed. Optimal couplings were ob tained during synthesis by using high-temperature couplings in conjunction with O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosph ate (HATU) activation. Improved purification schemes were developed that us e reverse- phase HPLC on a C1 column and elution with a 2:1 mixture of 1-pr opanol:1-butanol. Using these methods three very different transmembrane pe ptides all longer than 25 amino acids have been prepared: glycophorin-A, pr ion (110-137), and fibroblast growth factor receptor (368-397). (C) 1999 Ac ademic Press.