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Identification of the osteo‐inductive domain within the LMP‐3 protein (946.2)
Author(s) -
McMichael Ashley,
Zhou Liqiao,
Li Xeusen,
Robbins Paul
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.946.2
Subject(s) - microbiology and biotechnology , ectopic expression , transfection , fusion protein , chemistry , green fluorescent protein , gene , transduction (biophysics) , bone morphogenetic protein , reporter gene , context (archaeology) , biology , gene expression , recombinant dna , biochemistry , paleontology
LIM Mineralization protein (LMP‐3), a transcript variant form the original LMP protein has proven to be a novel osteogenic molecule when overexpressed, inducing expression of osteogenic factors include BMP‐2, BMP‐6 and Osterix (Osx). Expression of LMP‐3 by gene transfer results in ectopic bone formation in vivo. The goal of this project is to identify the minimal osteogenic domain in LMP‐3, which then could be delivered intracellularly using a protein transduction domain to induce osteogenesis for bone healing applications. To identify the minimal osteogenic domain, we generated a series of deletion constructs in LMP in the context of a LMP‐3‐eGFP fusion. MC3T3 cells were transfected with the different deletion mutants of LMP‐3 and the activation of an Osterix promoter‐luciferase reporter plasmid determined. Fragment F3 initially was shown to contain the osteoinductive domain of LMP‐3. Further deletion analysis of the F3 fragment was then performed. Within the F3 sequence, fragments F4 and F8 activated the OSX promoter. These results suggest that a small domain in LMP‐3 is sufficient to induce expression of osteoinductive genes required for bone formation and healing. Future studies will assess the functionality of these specific fragments in activating other osteogenic genes that lead to bone formation. Moreover, we will assess the ability of F4 and F8 to induce osteogenesis when delivered using a protein transduction domain. These LMP‐3 derived osteogenic peptides could be used to for bone fractures, bone defects, spine fusions, and invertebral disc regeneration.