Detection of protein-ligand interaction on the membranes using C-terminus biotin-tagged alamethicin.
C-terminal biotin-tagged alamethicin, which has several alpha-aminoisobutyric acid (Aib) residues in its sequence, was synthesized by the preparation of the protected peptide segment using the 2-chlorotrityl resin, followed by conjugation with biotin hydrazide. Suppression of the channel current of the biotin-tagged alamethicin by the addition of streptavidin to the electrolyte was monitorable in real time using the planar lipid-bilayer method. The system was also applicable to the detection of interaction of the biotin-tagged alamethicin with the anti-biotin antibody.
Zhang
Y
Y
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
Futaki
Shiroh
S
Kiwada
Tatsuto
T
Sugiura
Yukio
Y
D000408:Alamethicin / Q000138:chemical synthesis / Q000737:chemistry*; D000906:Antibodies / Q000737:chemistry; D001710:Biotin / Q000737:chemistry / Q000276:immunology; D015640:Ion Channel Gating / Q000187:drug effects; D007473:Ion Channels / Q000737:chemistry; D007476:Ionophores / Q000138:chemical synthesis / Q000737:chemistry*; D008024:Ligands; D008051:Lipid Bilayers / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D008954:Models, Biological; D011485:Protein Binding; D019809:Streptavidin / Q000737:chemistry
D000906:Antibodies; D007473:Ion Channels; D007476:Ionophores; D008024:Ligands; D008051:Lipid Bilayers; D008565:Membrane Proteins; D000408:Alamethicin; D001710:Biotin; D019809:Streptavidin