Sung, Jeong Soo Bong, Ji-Hong Yun, Tae Gyeong Han, Yeonju Park, Yusun Jung, Jaeyong Lee, Soo Jeong Kang, Min-Jung Jose, Joachim Lee, Misu Pyun, Jae-Chul 2024-01-19T12:00:57Z 2024-01-19T12:00:57Z 2022-06-30 2022-06 1043-1802 https://pubs.kist.re.kr/handle/201004/115134 Inhibitors for monoamine oxidase-B (MAO-B) were screened from an F-V library with a randomized complementarity-determining region 3 (CDR3) region using a monoclonal antibody against dopamine. As the first step, the F-V library was expressed on the outer membrane of E. coli by site-directed mutagenesis of the randomized CDR3 region. Among the F-V library, variants with a binding affinity to monoclonal antibodies against dopamine were screened and cloned. From the comparison of the binding activity of the screened clones to a control clone with a modified F-V antibody (only with CDR1 and CDR2), the CDR3 regions of screened clones were determined to directly interact with the monoclonal antibody against dopamine. These CDR3 sequences were then synthesized as mimotopes (mimicking peptides) of dopamine. The inhibitory activity of two mimotopes against MAO-B was analyzed using HeLa cells overexpressing MAO-B, as well as using activated human astrocytes; their inhibitory activity was compared to that of a commercial inhibitor of MAO-B, selegiline. The inhibition efficiency of the two mimotopes (in comparison with selegiline) was estimated to be 67.2% and 69.4% in the HeLa cells and 64.4% and 58.0% in the human astrocytes. The gene expression pattern in astrocytes after treatment with the two mimotopes was also analyzed and compared with that in the human astrocytes treated with selegiline. Finally, the interaction between two mimotopes and MAO-B was analyzed using docking simulation, and the candidate regions of MAO-B for the interaction with each mimotope were explored through the docking simulation. English AMER CHEMICAL SOC Antibody-Mediated Screening of Peptide Inhibitors for Monoamine Oxidase-B (MAO-B) from an Autodisplayed F-V Library Article 10.1021/acs.bioconjchem.2c00107 1 BIOCONJUGATE CHEMISTRY, v.33, no.6, pp.1166 - 1178 BIOCONJUGATE CHEMISTRY 33 6 1166 1178 N scie scopus 000812430500001 Biochemical Research Methods Biochemistry & Molecular Biology Chemistry, Multidisciplinary Chemistry, Organic Biochemistry & Molecular Biology Chemistry Article ESCHERICHIA-COLI OUTER-MEMBRANE DIAGNOSIS DIVERSITY PROTEINS STRESS REGION ASSAY