title: A novel Nogo-66 receptor antagonist peptide promotes neurite regeneration in vitro.
abstract: The Nogo-66 receptor (NgR1), a receptor for Nogo-A, contributes to the inhibition of axonal regeneration in the adult central nervous system after traumatic injuries. Thus, NgR1 has been considered a critical target in axon regeneration therapy. Here, we identified a specific NgR1 antagonist peptide (HIYTALV, named NAP2) which promotes neurite regeneration in vitro from a phage display heptapeptide library. NAP2 was co-localized with NgR1 on the surface of PC12 cells and cerebellar granule cells (CGCs) by immunofluorescence assay. Horseradish peroxidase (HRP)-streptavidin-biotin assay further showed that NAP2 binds to NgR1 and the dissociation constant (Kd) was 0.45 μM Functional analyses indicated that NAP2 could reduce the inhibitory effects of Nogo-66 on neurite outgrowth in differentiated PC12 cells and CGCs by blocking the Nogo-66-induced activation of Rho-associated coiled coil-containing protein kinase (ROCK), collapsin response mediator protein 2 (CRMP2) and myosin light chain (MLC). Taken together, the small molecule NgR1 antagonist peptide NAP2 (MW: 815.98Da) has a potential ability in crossing blood brain barrier and will be a promising therapeutic agent for the treatment of spinal cord injury and neurodegenerative diseases.
authors.affiliation: Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China.
authors.forename: Zhongqing
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, China.
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Dai
authors.affiliation: Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China.
authors.forename: Shuwen
authors.initials: S
authors.lastname: Jiang
authors.affiliation: Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, China.
authors.forename: Guofeng
authors.initials: G
authors.lastname: Lou
authors.affiliation: Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, China.
authors.forename: Qiaoyu
authors.initials: Q
authors.lastname: Cao
authors.affiliation: Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China.
authors.forename: Rendong
authors.initials: R
authors.lastname: Hu
authors.affiliation: Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, China.
authors.forename: Yadong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, China.
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Su
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau 999078, China.
authors.forename: Meiwan
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China.
authors.forename: Huanmin
authors.initials: H
authors.lastname: Luo
authors.affiliation: Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China; Guangzhou Key Laboratory of Environmental Exposure and Health, Jinan University, Guangzhou 510632, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Lin
authors.affiliation: Institue of Hydrobiology, Jinan University, Guangzhou 510632, China. Electronic address: tsunjun@jnu.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China; Guangzhou Key Laboratory of Environmental Exposure and Health, Jinan University, Guangzhou 510632, China. Electronic address: xiaofei@jnu.edu.cn.
authors.forename: Fei
authors.initials: F
authors.lastname: Xiao
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008024:Ligands; D009185:Myelin Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D018994:Myosin Light Chains / Q000378:metabolism; D009416:Nerve Regeneration; D009419:Nerve Tissue Proteins / Q000378:metabolism; D016501:Neurites / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D000070798:Nogo Proteins; D009842:Oligopeptides / Q000494:pharmacology*; D016716:PC12 Cells; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D009185:Myelin Proteins; D018994:Myosin Light Chains; D009419:Nerve Tissue Proteins; D000070798:Nogo Proteins; D009842:Oligopeptides; C095114:collapsin response mediator protein-2; D054460:rho-Associated Kinases
keywords: Antagonist peptide; Neurite regeneration; Nogo-66 receptor; Phage display