The receptor for advanced glycation end-products (RAGE) directly binds to ERK by a D-domain-like docking site.
The receptor for advanced glycation end-products (RAGE)-mediated cellular activation through the mitogen-activated protein kinase (MAPK) cascade, activation of NF-kappaB and Rho family small G-proteins, cdc42/Rac, is implicated in the pathogenesis of inflammatory disorders and tumor growth/metastasis. However, the precise molecular mechanisms for the initiation of cell signaling by RAGE remain to be elucidated. In this study, proteins which directly bind to the cytoplasmic C-terminus of RAGE were purified from rat lung extracts using an affinity chromatography technique and identified to be extracellular signal-regulated protein kinase-1 and -2 (ERK-1/2). Their interactions were confirmed by immunoprecipitation of ERK-1/2 from RAGE-expressing HT1080 cell extracts with anti-RAGE antibody. Furthermore, the augmentation of kinase activity of RAGE-bound ERK upon the stimulation of cells with amphoterin was demonstrated by determining the phosphorylation level of myelin basic protein, an ERK substrate. In vitro binding studies using a series of C-terminal deletion mutants of human RAGE revealed the importance of the membrane-proximal cytoplasmic region of RAGE for the direct ERK-RAGE interaction. This region contained a sequence similar to the D-domain, a ERK docking site which is conserved in some ERK substrates including MAPK-interacting kinase-1/2, mitogen- and stress-activated protein kinase-1, and ribosomal S6 kinase. These data suggest that ERK may play a role in RAGE signaling through direct interaction with RAGE.
Ishihara
Katsuya
K
Discovery Biology, Tsukuba Research Institute, Novartis Pharma K.K., Okubo 8, Tsukuba-shi, Ibaraki 300-2611, Japan.
Tsutsumi
Kae
K
Kawane
Shiho
S
Nakajima
Motowo
M
Kasaoka
Tatsuhiko
T
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D002457:Cell Extracts; D002478:Cells, Cultured; D017124:Conserved Sequence; D003593:Cytoplasm / Q000378:metabolism; D024243:HMGB1 Protein / Q000494:pharmacology; D006801:Humans; D008168:Lung / Q000737:chemistry / Q000378:metabolism; D008297:Male; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000302:isolation & purification / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000302:isolation & purification / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion
D002457:Cell Extracts; D024243:HMGB1 Protein; D004676:Myelin Basic Protein; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases