title: Proteomic analysis of cPKCβII-interacting proteins involved in HPC-induced neuroprotection against cerebral ischemia of mice.
abstract: Hypoxic preconditioning (HPC) initiates intracellular signaling pathway to provide protection against subsequent cerebral ischemic injuries, and its mechanism may provide molecular targets for therapy in stroke. According to our study of conventional protein kinase C βII (cPKCβII) activation in HPC, the role of cPKCβII in HPC-induced neuroprotection and its interacting proteins were determined in this study. The autohypoxia-induced HPC and middle cerebral artery occlusion (MCAO)-induced cerebral ischemia mouse models were prepared as reported. We found that HPC reduced 6 h MCAO-induced neurological deficits, infarct volume, edema ratio and cell apoptosis in peri-infarct region (penumbra), but cPKCβII inhibitors Go6983 and LY333531 blocked HPC-induced neuroprotection. Proteomic analysis revealed that the expression of four proteins in cytosol and eight proteins in particulate fraction changed significantly among 49 identified cPKCβII-interacting proteins in cortex of HPC mice. In addition, HPC could inhibit the decrease of phosphorylated collapsin response mediator protein-2 (CRMP-2) level and increase of CRMP-2 breakdown product. TAT-CRMP-2 peptide, which prevents the cleavage of endogenous CRMP-2, could inhibit CRMP-2 dephosphorylation and proteolysis as well as the infarct volume of 6 h MCAO mice. This study is the first to report multiple cPKCβII-interacting proteins in HPC mouse brain and the role of cPKCβII-CRMP-2 in HPC-induced neuroprotection against early stages of ischemic injuries in mice.
authors.affiliation: Department of Neurobiology and Beijing Institute for Neuroscience, Capital Medical University, Beijing, China.
authors.forename: Xiangning
authors.initials: X
authors.lastname: Bu
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
authors.forename: Xuan
authors.initials: X
authors.lastname: Yang
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Liu
authors.forename: Jianli
authors.initials: J
authors.lastname: Du
authors.forename: Jing
authors.initials: J
authors.lastname: Liang
authors.forename: Qunyuan
authors.initials: Q
authors.lastname: Xu
authors.forename: Junfa
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D001929:Brain Edema / Q000209:etiology / Q000517:prevention & control; D020520:Brain Infarction / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*; D002545:Brain Ischemia / Q000150:complications / Q000378:metabolism*; D030562:Databases, Protein; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D047468:Immunoprecipitation / Q000379:methods; D020287:In Situ Nick-End Labeling / Q000379:methods; D007211:Indoles / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D019194:Ischemic Preconditioning / Q000379:methods*; D008297:Male; D008301:Maleimides / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009460:Neurologic Examination / Q000379:methods; D018696:Neuroprotective Agents / Q000627:therapeutic use; D010455:Peptides / Q000627:therapeutic use; D010766:Phosphorylation / Q000187:drug effects / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism*; D064546:Protein Kinase C beta; D040901:Proteomics / Q000379:methods*; D016622:Silver Staining; D013997:Time Factors
chemical_list: C465664:2-(1-(3-dimethylaminopropyl)-5-methoxyindol-3-yl)-3-(1H-indol-3-yl)maleimide; D004791:Enzyme Inhibitors; D007211:Indoles; D036341:Intercellular Signaling Peptides and Proteins; D008301:Maleimides; D009419:Nerve Tissue Proteins; D018696:Neuroprotective Agents; D010455:Peptides; C095114:collapsin response mediator protein-2; C099154:ruboxistaurin; D011493:Protein Kinase C; D064546:Protein Kinase C beta