RRC ID 73302
著者 Sugimoto H, Fukuda S, Yokawa S, Hori M, Ninomiya H, Sato T, Miyazawa K, Kawai T, Furuno T, Inouye S, Goto S, Suzuki T.
タイトル Visualization of osteocalcin and bone morphogenetic protein 2 (BMP2) secretion from osteoblastic cells by bioluminescence imaging.
ジャーナル Biochem Biophys Res Commun
Abstract The secretions of osteocalcin and bone morphogenetic protein 2 (BMP2) from living osteoblastic cells were visualized for the first time using a method of video-rate bioluminescence imaging. The fusion proteins with Gaussia luciferase (GLase) for mouse osteocalcin and BMP2 (OC-GLase and BMP2-GLase, respectively) expressed in osteoblastic MC3T3-E1 cells were correctly processed and secreted. In the video images of exocytotic secretion, the luminescence spots of OC-GLase and BMP2-GLase disappeared rapidly and gradually, respectively, indicating different manners of these proteins in diffusion. Notably, a deletion mutant of BMP2 (Δ3BMP2-GLase) lacking three basic amino acid residues in the N-terminal region for binding to heparan sulfate showed rapidly disappearing luminescence spots. In our imaging conditions, the half-life of luminescence for the spots of Δ3BMP2-GLase (1.61 ± 0.20 s) was similar to that of OC-GLase (1.22 ± 0.14 s) but not to that of BMP2-GLase (4.31 ± 0.41 s). These results suggest that, in contrast to osteocalcin, the diffusion of BMP2 from cells occurred slowly after exocytosis. Thus, our bioluminescence imaging method is useful to study the diffusion properties of secreted proteins in exocytosis.
巻・号 635
ページ 203-209
公開日 2022-12-20
DOI 10.1016/j.bbrc.2022.10.042
PII S0006-291X(22)01436-X
PMID 36279682
MeSH Animals Bone Morphogenetic Protein 2* / metabolism Cell Communication* Cell Differentiation Cell Line Luciferases / genetics Luciferases / metabolism Mice Osteoblasts / metabolism Osteocalcin / genetics Osteocalcin / metabolism
IF 2.985
リソース情報
ヒト・動物細胞 MC3T3-E1(RCB1126)