Details

Autor(en) / Beteiligte

• Manandhar, Yasodha
• Wang, Wei
• Inoue, Jin
• Hayashi, Nobuhiro
• Uzawa, Takanori
• Ito, Yutaka
• Aigaki, Toshiro
• Ito, Yoshihiro

Titel

Interactions of in vitro selected fluorogenic peptide aptamers with calmodulin

Ist Teil von

• Biotechnology letters, 2017-03, Vol.39 (3), p.375-382

Ort / Verlag

Dordrecht: Springer Netherlands

Erscheinungsjahr

2017

Quelle

MEDLINE

Beschreibungen/Notizen

• Objectives We examined the importance of aptamer usage under the same condition as the selection process by employing the previously selected aptamers for calmodulin (CaM) which includes a non-natural fluorogenic amino acid, 7-nitro-2,1,3-benzoxadiazole. Results We added five amino acids at the N -terminus which was employed for the selection and then we tested the affinity and selectivity for CaM binding. Surface plasmon resonance and fluorescence measurements showed that the additional amino acids for one of the aptamers drastically improved binding affinity to CaM, indicating the importance of aptamer use under the same conditions as the selection process. Such drastic improvement in affinity was not observed for the sequence which had been reported previously. Nuclear magnetic resonance data identified that the primary binding site is located in a C -terminal of CaM and the additional residues enhance interactions with CaM. Conclusions We found that the addition of the common sequence, which was employed for ribosome display, makes the affinity of a selected peptide as strong as the previously reported peptide.