C-Mannosylated peptides derived from the thrombospondin type 1 repeat interact with Hsc70 to modulate its signaling in RAW264.7 cells
| Autor: | Midori Ikezaki, Shino Manabe, Yuriko Ohta, Yoshito Ihara, Eiji Muroi, Yukishige Ito, Yoko Inai, In Sook Lee Matsui |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Conformational change
Cell signaling animal structures Blotting Western Fluorescence Polarization macromolecular substances Biochemistry Thrombospondin 1 Mice Animals RNA Messenger Gene Cells Cultured Thrombospondin Reverse Transcriptase Polymerase Chain Reaction Tumor Necrosis Factor-alpha Chemistry Kinase Macrophages HSC70 Heat-Shock Proteins Tryptophan SUPERFAMILY MAP Kinase Kinase Kinases Peptide Fragments Cell biology carbohydrates (lipids) Microscopy Fluorescence Functional module Tumor necrosis factor alpha Mannose Signal Transduction |
| Zdroj: | Glycobiology. 20:1298-1310 |
| ISSN: | 1460-2423 0959-6658 |
| Popis: | The thrombospondin type 1 repeat (TSR) is a functional module of proteins called TSR superfamily proteins (e.g., thrombospondin, F-spondin, mindin, etc.) and includes a conserved Trp-x-x-Trp (W-x-x-W) motif, in which the first Trp residue is preferably modified by C-mannosylation. We previously reported that synthesized C-mannosylated TSR-derived peptides (e.g., C-Man-WSPW) specifically enhanced lipopolysaccharide-induced signaling in macrophage-like RAW264.7 cells. In this study, we searched for the proteins that bind to C-mannosylated TSR-derived peptides in RAW264.7 cells and identified heat shock cognate protein 70 (Hsc70). The binding affinity of Hsc70 for C-mannosylated peptides in solution was higher than that for the peptides without C-mannose. The binding was influenced by a nucleotide-induced conformational change of Hsc70, and C-mannosylated peptides preferred the substrate-binding domain of Hsc70. Furthermore, in RAW264.7 cells, addition of Hsc70 stimulated cellular signaling to produce tumor necrosis factor-alpha, via transforming growth factor-beta-activated kinase 1, and the Hsc70-induced signaling was enhanced more in the presence of the peptides with C-mannose than that without C-mannose, suggesting functional interaction between Hsc70 and the C-mannosylated peptides in the cells. Together, these results demonstrate a novel function of the C-mannosylation of TSR-derived peptides in terms of interaction with Hsc70 to regulate cellular signaling. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|