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Design and utility of CCN2 anchor peptide aptamers
Authors:Harumi Kawaki  Satoshi Kubota  Eriko Aoyama  Naoya Fujita  Hiroshi Hanagata  Akira Miyauchi  Kenta Nakai  Masaharu Takigawa
Institution:1. Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan;2. Biodental Research Center, Okayama University Dental School, Okayama, Japan;3. Human Genome Center, Institute of Medical Science, University of Tokyo, Tokyo, Japan;4. Research Laboratory, Higeta Shoyu Co. Ltd., Choshi, Japan;5. Kazusa Laboratory, Protein Express Co. Ltd., Kisarazu, Japan
Abstract:CCN family protein 2/connective tissue growth factor (CCN2/CTGF) consists of 4 conserved modules that are highly interactive with a number of biomolecules. With such interaction, CCN2 exerts multiple functions by forming an extracellular information network. In the present study, we screened for dodecapeptide sequences that bound to each module of human CCN2 by using a bacteriophage display library. Thereafter, consensus amino acid sequences for the binding to individual modules were extracted in silico and utilized to design anchor peptide aptamers that would facilitate the interaction between CCN2 and other molecules. Direct binding of a few peptides to CCN2 was confirmed by surface plasmon resonance analysis. Subsequent biological assay indicated that one such peptide was capable of promoting the proliferation of CCN2-producing chondrocytic cells. This cell biological activity was found to be sequence specific and CCN2 dependent. Since CCN2/CTGF was shown to be effective in articular cartilage/bone regeneration in vivo, utility of such peptide aptamers in CCN2-associated regenerative therapeutics is suggested herein.
Keywords:CCN family  CTGF  Aptamer  Chondrocyte  Tissue regeneration
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