Spontaneous integration of transmembrane peptides into a bacterial magnetic particle membrane and its application to display of useful proteins.
An antimicrobial peptide, temporin L, and its derivative (TL-A2) were employed as anchor peptides and displayed streptavidin on a bacterial magnetic particle (BMP) membrane. The ribotoxin L3 loop (L3) and the arginine-chain peptide (R(12)), which are carrier peptides permeable to eukaryotic cell membranes, were also used. The peptides were labeled with a fluorescent dye, 4-fluoro-7-nitrobenzofurazan (NBD), at the N-terminal region (NBD-peptides) and mixed with BMPs. A specific integration of NBD-temporin L into a BMP membrane was observed. The basic amino acids in temporin L played an important role in the integration into BMPs. Biotin conjugated to the N-terminus of temporin L was integrated into a BMP membrane. The C-terminus of temporin L was incorporated into a BMP membrane, and the N-terminus was located on the BMP membrane surface. The present study shows that temporin L is a stable molecular anchor on BMPs by the binding of soluble protein to the N-terminus.
Tanaka
Tsuyoshi
T
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-Cho, Koganei, Tokyo, Japan.
Takeda
Hajime
H
Kokuryu
Yoriko
Y
Matsunaga
Tadashi
T
D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008280:Magnetics; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000737:chemistry / Q000502:physiology*; D008567:Membranes, Artificial; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012965:Sodium Chloride / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry; D013499:Surface Properties
D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins; D008565:Membrane Proteins; D008567:Membranes, Artificial; D010446:Peptide Fragments; D010455:Peptides; C469676:temporin L, Rana temporaria; D012965:Sodium Chloride; D001710:Biotin; D019809:Streptavidin