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 insoluble transmembrane peptides has been developed. Optimal couplings were obtained during synthesis by using high-temperature couplings in conjunction with O-(7-azabenzotriazol-1-yl)-1,1,3, 3-tetramethyluronium hexafluorophosphate (HATU) activation. Improved purification schemes were developed that use reverse- phase HPLC on a C1 column and elution with a 2:1 mixture of 1-propanol:1-butanol. Using these methods three very different transmembrane peptides all longer than 25 amino acids have been prepared: glycophorin-A, prion (110-137), and fibroblast growth factor receptor (368-397).
Glover
K J
KJ
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California 92093-0601, USA.
Martini
P M
PM
Vold
R R
RR
Komives
E A
EA
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006021:Glycophorins / Q000138:chemical synthesis* / Q000737:chemistry; D007202:Indicators and Reagents; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry; D011328:Prions / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D017468:Receptors, Fibroblast Growth Factor / Q000737:chemistry*
D006021:Glycophorins; D007202:Indicators and Reagents; D010446:Peptide Fragments; D011328:Prions; D017468:Receptors, Fibroblast Growth Factor