Heterologous expression of BACE1 and its interaction with Codonopsis pilosula polysaccharides and Lobetyolin.

Autor: Zhao F; School of life science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China., Fan L; School of life science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China., Yang J; Lanzhou University Second Hospital, Lanzhou 730000, China., Yang M; School of life science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China., Zhang C; Lanzhou University of Technology Hospital, Lanzhou 730000, China., Wang F; School of life science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China. Electronic address: wangfang@lut.edu.cn., Wang Y; School of life science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China. Electronic address: wangyg@lut.edu.cn.
Jazyk: angličtina
Zdroj: International journal of biological macromolecules [Int J Biol Macromol] 2024 Oct; Vol. 277 (Pt 2), pp. 133440. Date of Electronic Publication: 2024 Jun 28.
DOI: 10.1016/j.ijbiomac.2024.133440
Abstrakt: BACE1, a crucial enzyme in the amyloid-β deposition theory of Alzheimer's disease (AD), is targeted by Codonopsis pilosula, a traditional tonic believed to impede AD onset. However, the specific active compounds responsible for its effects remain elusive. Our prior network pharmacology research identified C. pilosula polysaccharides (CPPS) and Lobetyolin may serve as potential inhibitors of AD by suppressing amyloidogenesis. Here, we recombinantly expressed BACE1 under varied conditions and assessed its activity using Fluorescence Resonance Energy Transfer technology. Through spectroscopy, molecular docking, and dynamics, we elucidated the interactions of CPPS, Lobetyolin, and BACE1. Optimal BACE1 expression occurred at 22 °C with 0.4 mM IPTG for 6 h, yielding a 72 kDa protein. Enzyme kinetics displayed a maximum rate of 4096 μmol/min and a Michaelis constant of 16 mg/mL for BACE1. Spectroscopic analysis revealed differing binding affinities of the compounds at various temperatures, peaking at 293 K. Lobetyolin exhibited superior binding to BACE1 compared to CPPS, driven by hydrophobic and electrostatic forces. Molecular docking and dynamics highlighted hydrophobic amino acids' role in BACE1 interactions with Lobetyolin and CPPS, with binding energy < -1.2 kcal/mol signifying strong affinities. Notably, Lobetyolin and CPPS showed higher BACE1 affinity than APP, with the Lobetyolin-BACE1 complex being the most stable.
Competing Interests: Declaration of competing interest Declaration of competing interest the authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
Externí odkaz: