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.