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A technology of mutants and receptors, applied in the field of NRG mutants
Active Publication Date: 2006-01-04
ZENSUN (SHANGHAI) SCI & TECH CO LTD
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The study found that rhNRG activates ErbB4 receptors to treat heart disease while activating other ErbB receptors to produce side effects
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[0069] Example Screening of NRG mutants that specifically activate ErbB receptors
[0070] 1. Method
[0071] The EGF-like domain of NRG-1β can bind and activate the receptor to perform biological functions. In this study, in order to simplify the model, the EGF-like domain of NRG-1β (NRG-β 177-229 , residues 1-52), hereinafter referred to as NRG-1β. Due to the lack of three-dimensional structure information of NRG-1β and receptor ErbB, it is necessary to use theoretical prediction methods to construct their three-dimensional protein structures, so as to study their ligand / receptor interactions at the microscopic level. In addition, so far, there have been many successful examples of using homology modeling, molecular dynamics simulation and free energy calculations to study protein / small molecule ligands or protein / protein interactions at the atomic level in theoretical biology research. 12-17 . The homologous protein modeling method can accurately construct the three-dim...
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Abstract
The present invention provides a method of screening NRG mutant for activating ErbB acceptor specifically. The method includes the following steps: establishing 3D model of complex of NRG, ErbB3, ErbB4, NRG / ErbB3 and NRG / ErbB4 in a homogeneous model establishing process; molecular kinetically simulating conformation and stability of NRG / ErbB3 and NRG / ErbB4; MM / PBSA process of calculating the binding free energy between NRG and ErbB3 or ErbB4; scanning and calculating process based on alanine theory to determine the NRG and acceptor affinity change during the mutation from NRG residue to alanine and determining NRG mutant for activating ErbB acceptor specifically. The present invention also provides the NRG mutant and its application and preparation process.
Description
Technical field: [0001] The invention relates to an NRG mutant that specifically activates ErbB receptors, a screening method and applications thereof. Background technique: [0002] The role of Neuregulins (NRGs) and receptor ErbBs family in cell signal transduction in the formation of nerve, muscle, epithelial and other tissues and organs has been widely known (Lemke, Mol. Cell. Neurosci. 7: 247-262 , 1996; Burden et al., Neuron 18:847-855, 1997). [0003] The more in-depth study is the protein encoded by the NRG-1 gene, which includes about 15 different subtypes with the same structure (Lemke, Mol. Cell. Neurosci. 7: 247-262, 1996 and Peles and Yarden, BioEssays 15: 815- 824, 1993). NRG-1 subtypes include Neu Differentiation Factor (NDF; Peles et al., Cell 69, 205-216, 1992 and Wen et al., Cell 69, 559-572, 1992), Heregulin (HRG; Holmes et al., Science 256: 1205-1210, 1992), Acetylcholine Receptor Inducing Activity (ARIA; Falls et al., Cell 72: 801-815, 1993), and glia...
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