Directed evolution of angiotensin II-inhibiting peptides using a microbead display.
Angiotensin II (ang II), an octapeptide (DRVYVHPF), can regulate blood pressure by binding specifically to its receptor, AT1. A peptide (VVIVIY) in the first transmembrane of AT1 has been found, via peptide array technology, to have an affinity for ang II. In this study, the peptide P2, which contained the VVIVIY sequence, was mutated and screened using microbead display technology that utilized emulsion PCR and cell-free protein synthesis. After one round of screening, the binding activities of collected mutants were estimated using flow cytometry and a peptide array. Two of these exhibited improved association rate constants to ang II, compared to the P2 peptide.
Gan
Rui
R
Laboratory of Molecular Biotechnology, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
Furuzawa
Seiji
S
Kojima
Takaaki
T
Kanie
Kei
K
Kato
Ryuji
R
Okochi
Mina
M
Honda
Hiroyuki
H
Nakano
Hideo
H
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D019020:Directed Molecular Evolution / Q000379:methods*; D066298:In Vitro Techniques; D007700:Kinetics; D008863:Microspheres; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
D015342:DNA Probes; D009842:Oligopeptides; D019151:Peptide Library; D011994:Recombinant Proteins; D000804:Angiotensin II