title: Derivation of Self-inhibitory Helical Peptides to Target Rho-kinase Dimerization in Cerebrovascular Malformation: Structural Bioinformatics Analysis and Peptide Binding Assay.
abstract: 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 KD 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.
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Xuyang
authors: X
authors: Wang
authors: Department of Neurosurgery, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, 200080, China.
authors: Dianqi
authors: D
authors: Hou
authors: The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, China.
authors: Weiwei
authors: W
authors: Dai
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Wenwei
authors: W
authors: Gao
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Shiming
authors: S
authors: Ju
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Heli
authors: H
authors: Cao
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Lin
authors: L
authors: Zhang
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Gan
authors: G
authors: Wang
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Yan
authors: Y
authors: Guo
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China.
authors: Shiwen
authors: S
authors: Chen
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai, 200233, China. tianhengli@126.com, tianhengli1964@yahoo.com.cn.
authors: Hengli
authors: H
authors: Tian
authors: Department of Neurosurgery, Shanghai Jiao Tong University Affiliated 6th People's Hospital South Campus, Shanghai, 201400, China. lzq_999@163.com.
authors: Zhiqiang
authors: Z
authors: Li
mesh_terms: D004353:Drug Evaluation, Preclinical; D005454:Fluorescence Polarization; D002538:Intracranial Arteriovenous Malformations / Q000188:drug therapy / Q000201:enzymology; D056004:Molecular Dynamics Simulation; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D054730:Protein Interaction Domains and Motifs; D047428:Protein Kinase Inhibitors / Q000737:chemistry*; D055503:Protein Multimerization; D013816:Thermodynamics; D054460:rho-Associated Kinases / Q000737:chemistry*
chemical_list: D010455:Peptides; D047428:Protein Kinase Inhibitors; D054460:rho-Associated Kinases
keywords: Rho-kinase; cerebrovascular malformation; dimerization; self-inhibitory peptide