Directed evolution of angiotensin II-inhibiting peptides using a microbead display

作者：Gan Rui; Furuzawa Seiji; Kojima Takaaki; Kanie Kei; Kato Ryuji; Okochi Mina; Honda Hiroyuki; Nakano Hideo^*

来源：Journal of Bioscience and Bioengineering, 2010, 109(4): 411-417.

DOI：10.1016/j.jbiosc.2009.10.009

• Single-molecule PCR
• Microbead display
• Cell-free protein synthesis
• Directed evolution
• Angiotensin II

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.

• 出版日期2010-4