Application of a Fluorescent Biosensor in Determining the Binding of 5-HT to Calmodulin
Por:
Vasquez-Bochm, L. X., Velazquez-Lopez, Isabel, Mata, Rachel, Sosa-Peinado, Alejandro, Cano-Sanchez, Patricia, Gonzalez-Andrade, Martin
Publicada:
1 sep 2021
Resumen:
Here, we show the utility of the fluorescent biosensor hCaM-M124C-mBBr
in detecting and determining the affinity of serotonin (5-HT). We
obtained a K-d of 5-HT (0.71 mu m) for the first time, the same order of
magnitude as most anti-CaM drugs. This data can contribute to
understanding the direct and indirect modulation of CaM on its binding
proteins when the 5-HT concentration varies in different tissues or
explain some of the side effects of anti-CaM drugs. On the other hand,
molecular modeling tools help the rational design of biosensors and
adequately complement the experimental results. For example, the docking
study indicates that 5-HT binds at the same site as chlorpromazine (site
1) with a theoretical K-i of 2.84 mu M; while the molecular dynamics
simulations indicate a stability of the CaM-5-HT complex with a
theoretical Delta G of -4.85 kcal mol(-1), where the enthalpy
contribution is greater. Thus, the combination of biotechnology and
bioinformatics helps in the design and construction of more robust
biosensors.
Filiaciones:
Vasquez-Bochm, L. X.:
Facultad de Química, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico
Velazquez-Lopez, Isabel:
Laboratorio de Biosensores y Modelaje Molecular, Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico
Gonzalez-Andrade, M (Corresponding Author), Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Lab Biosensores & Modelaje Mol, Ciudad De Mexico 04510, Mexico. Vasquez-Bochm, L. X
Mata, Rachel:
Facultad de Química, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico
Gonzalez-Andrade, M (Corresponding Author), Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Lab Biosensores & Modelaje Mol, Ciudad De Mexico 04510, Mexico. Vasquez-Bochm, L. X
Sosa-Peinado, Alejandro:
Laboratorio de Biosensores y Modelaje Molecular, Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico
Gonzalez-Andrade, M (Corresponding Author), Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Lab Biosensores & Modelaje Mol, Ciudad De Mexico 04510, Mexico. Vasquez-Bochm, L. X
Cano-Sanchez, Patricia:
Instituto de Química, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico
Gonzalez-Andrade, M (Corresponding Author), Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Lab Biosensores & Modelaje Mol, Ciudad De Mexico 04510, Mexico. Vasquez-Bochm, L. X
Gonzalez-Andrade, Martin:
Laboratorio de Biosensores y Modelaje Molecular, Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Ciudad de México, 04510, Mexico
Gonzalez-Andrade, M (Corresponding Author), Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Lab Biosensores & Modelaje Mol, Ciudad De Mexico 04510, Mexico. Vasquez-Bochm, L. X
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