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Derivation of Self‐inhibitory Helical Peptides to Target Rho‐kinase Dimerization in Cerebrovascular Malformation: Structural Bioinformatics Analysis and Peptide Binding Assay
Author(s) -
Wang Xuyang,
Hou Dianqi,
Dai Weiwei,
Gao Wenwei,
Ju Shiming,
Cao Heli,
Zhang Lin,
Wang Gan,
Guo Yan,
Chen Shiwen,
Tian Hengli,
Li Zhiqiang
Publication year - 2016
Publication title -
molecular informatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.481
H-Index - 68
eISSN - 1868-1751
pISSN - 1868-1743
DOI - 10.1002/minf.201501022
Subject(s) - in silico , peptide , kinase , chemistry , molecular mechanics , rho associated protein kinase , biochemistry , peptidomimetic , protein kinase a , molecular dynamics , protein kinase domain , biophysics , biology , computational chemistry , gene , mutant
Rho‐kinase dimerization is essential for its kinase activity and biological function; disruption of the dimerization has recently been established as a new and promising therapeutics strategy for cerebrovascular malformation (CM). Based on Rho‐kinase dimer crystal structure we herein combined in silico analysis and in vitro assay to rationally derive self‐inhibitory peptides from the dimerization interface. Three peptides namely Hlp1 , Hlp2 and Hlp3 were successfully designed that have potential capability to rebind at the dimerization domain of Rho‐kinase. Molecular dynamics (MD) simulations revealed that these peptides are helically structured when bound to Rho‐kinase, but exhibit partially intrinsic disorder in unbound state. Binding free energy (BFE) analysis suggested that the peptides have a satisfactory energetic profile to interact with Rho‐kinase. The computational findings were then substantiated by fluorescence anisotropy assays, conforming that the helical peptides can bind tightly to Rho‐kinase with affinity K D at micromolar level. These designed peptides are considered as lead molecular entities that can be further modified and optimized to obtain more potent peptidomimetics as self‐competitors to disrupt Rho‐kinase dimerization in CM.