Tyrosine phosphorylation of clathrin heavy chain under oxidative stress.
In mouse pancreatic insulin-producing betaTC cells, oxidative stress due to H(2)O(2) causes tyrosine phosphorylation in various proteins. To identify proteins bearing phosphotyrosine under stress, the proteins were affinity purified using an anti-phosphotyrosine antibody-conjugated agarose column. A protein of 180kDa was identified as clathrin heavy chain (CHC) by electrophoresis and mass spectrometry. Immunoprecipitated CHC showed tyrosine phosphorylation upon H(2)O(2) treatment and the phosphorylation was suppressed by the Src kinase inhibitor, PP2. The phosphorylation status of CHC affected the intracellular localization of CHC and the clathrin-dependent endocytosis of transferrin under oxidative stress. In conclusion, CHC is a protein that is phosphorylated at tyrosine by H(2)O(2) and this phosphorylation status is implicated in the intracellular localization and functions of CHC under oxidative stress. The present study demonstrates that oxidative stress affects intracellular vesicular trafficking via the alteration of clathrin-dependent vesicular trafficking.
Ihara
Yoshito
Y
Department of Biochemistry and Molecular Biology in Disease, Atomic Bomb Disease Institute, Nagasaki University School of Medicine, 1-12-4 Sakamoto, Japan.
Yasuoka
Chie
C
Kageyama
Kan
K
Wada
Yoshinao
Y
Kondo
Takahito
T
D000818:Animals; D002478:Cells, Cultured; D033922:Clathrin Heavy Chains / Q000378:metabolism*; D004705:Endocytosis / Q000502:physiology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D010766:Phosphorylation; D021381:Protein Transport / Q000502:physiology*; D011743:Pyrimidines / Q000494:pharmacology; D014168:Transferrin / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*; D019061:src-Family Kinases / Q000037:antagonists & inhibitors
C412373:AG 1879; D016877:Oxidants; D011743:Pyrimidines; D014168:Transferrin; D033922:Clathrin Heavy Chains; D014443:Tyrosine; D006861:Hydrogen Peroxide; D019061:src-Family Kinases