title: [Effect of zoledronate on protein interaction between Ca(2+)/calmodulin-dependent protein kinaseⅡ and calmodulin and expression of downstream genes during osteoclast differentiation].
abstract: Objective:To investigate the effect of zoledronate on protein interaction between Ca(2+)/calmodulin-dependent protein kinaseⅡ(CaMKⅡ) and calmodulin and protein expression of nuclear factor of activation of T cells-1 (NFATc1) and tartrate resistant acid phosphatase (TRAP) during osteoclast differentiation.Methods:Mouse RAW264.7 cells were divided into group A and B and were cultured. Group A was induced with 50 mg/L receptor activator of NF-κB ligand (RANKL) for osteoclastogenesis, and group B was treated with 1×10(-6) zoledronate for two days from day 2. Co-immunoprcipitation (Co-IP) and reverse Co-IP were used to detect the protein-binding between CaMKⅡ and calmodulin. Western-blotting and immunofluorescent cytochemistry were also used to detect the protein level of NFATc1 and TRAP in both groups. Osteoclast formation was also analyzed.Results:In group B, the number of osteoclasts, number and size of dentin resorption lacunaes were 11.3±1.5, 8.7±2.1 and (5 034.4±775.4) μm(2) respevtively, which were significantly lower than those (37.7±5.7, 23.0±4.0 and [15 042.7±1 906.0] μm(2)) in group A (P<0.01). Co-IP and reverse Co-IP examination indicated that protein-binding between CaMKⅡ and calmodulin significantly decreased by 59.8% and 50.9% in group B compared with group A (P<0.01). The protein level of calmodulin and CaMKⅡ in total cellular proteins also significantly decreased by 52.1% and 51.5% in group B compared with group A (P<0.01). NFATc1 and TRAP protein decreased by 52.4% and 38.9% in group B than in group A (P<0.01), respectively.Conclusions:Zoledronate could significantly inhibit protein-binding between CaMKⅡ and calmodulin and down-regulate protein level of NFATc1 and TRAP.
authors.affiliation: Department of Oral and Maxillofacial Surgery, College of Stomatology, North China University of Science and Technology, Tangshan Hebei 063000, China.
authors.forename: H
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Oral and Maxillofacial Surgery, College of Stomatology, North China University of Science and Technology, Tangshan Hebei 063000, China.
authors.forename: W
authors.initials: W
authors.lastname: Dong
authors.affiliation: Department of Oral and Maxillofacial Surgery, College of Stomatology, North China University of Science and Technology, Tangshan Hebei 063000, China.
authors.forename: M C
authors.initials: MC
authors.lastname: Qi
authors.affiliation: Department of Basic Medical Sciences, Department of Pathology, North China University of Science and Technology, Tangshan Hebei 063000, China.
authors.forename: H
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Oral and Maxillofacial Surgery, College of Stomatology, North China University of Science and Technology, Tangshan Hebei 063000, China.
authors.forename: X J
authors.initials: XJ
authors.lastname: Feng
authors.affiliation: Department of Oral and Maxillofacial Surgery, College of Stomatology, North China University of Science and Technology, Tangshan Hebei 063000, China.
authors.forename: L M
authors.initials: LM
authors.lastname: Wen
mesh_terms: D000818:Animals; D050071:Bone Density Conservation Agents / Q000494:pharmacology*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D004164:Diphosphonates / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D007093:Imidazoles / Q000494:pharmacology*; D051379:Mice; D050778:NFATC Transcription Factors / Q000378:metabolism; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D000071681:Tartrate-Resistant Acid Phosphatase / Q000378:metabolism*; D000077211:Zoledronic Acid
chemical_list: D050071:Bone Density Conservation Agents; D002147:Calmodulin; D004164:Diphosphonates; D007093:Imidazoles; D050778:NFATC Transcription Factors; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D000077211:Zoledronic Acid; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D000071681:Tartrate-Resistant Acid Phosphatase
keywords: Calcium-calmodulin-dependent protein kinase type 2; Calmodulin; Immunoprecipitation; Zolderonate