Developing sensor-proteins to detect CAPN3 protease activity

2014 Fiscal Year Final Research Report

• Project/Area Number: 23500477
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurophysiology and muscle physiology
• Research Institution: National Agriculture and Food Research Organization
• Principal Investigator: OJIMA Koichi 独立行政法人農業・食品産業技術総合研究機構, 畜産草地研究所 畜産物研究領域, 主任研究員 (60415544)
• Co-Investigator(Renkei-kenkyūsha): HATA Shoji (財)東京都医学研究機構, 東京都臨床医学総合研究所, 主席研究員 (10392375) ; ONO Yasuko (財)東京都医学研究機構, 東京都臨床医学総合研究所, 主席研究員 (20392376)
• Project Period (FY): 2011-04-28 – 2015-03-31
• Keywords: 骨格筋 / カルパイン

Outline of Final Research Achievements

Calpain-3 (CAPN3) is predominantly expressed in skeletal muscle and is Ca2+-requiring intracellular cysteine protease. In this study, we made sensor-proteins to detect CAPN3 protease activity using a fluorescence resonance energy transfer technique and examined whether CAPN3 protease activity was controlled by phosphorylation. Unfortunately, it was very tough to detect protease activity of CAPN3 in living muscle cells although sensor-proteins were expressed well and were cleaved by CAPN3 in vitro experiments. On the other hand, we showed that rapid and exhaustive autolysis of CAPN3 was slightly attenuated by the defect of phosphorylation of Ser residue at 629 of CAPN3.

Free Research Field

骨格筋