Imaging and photoregulation of cellular functions by using functional molecule-protein hybrid probes

2017 Fiscal Year Final Research Report

• Project/Area Number: 15H03120
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Chemical biology
• Research Institution: Tohoku University
• Principal Investigator: MIZUKAMI Shin 東北大学, 多元物質科学研究所, 教授 (30420433)
• Co-Investigator(Renkei-kenkyūsha): KIKUCHI KAZUYA 大阪大学, 大学院工学研究科, 教授 (70292951)
• Research Collaborator: MATSUI YUSUKE 大阪大学, 大学院工学研究科, 大学院生 ; SATO RYOTA 大阪大学, 大学院工学研究科, 大学院生 ; AKAZAWA KAZUKI 大阪大学, 大学院工学研究科, 大学院生 ; SUZUKI SHUNSUKE 大阪大学, 大学院工学研究科, 大学院生 ; IMOTO TAKUMA 大阪大学, 大学院工学研究科, 大学院生
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 蛍光イメージング / タンパク質ラベル化 / Mg2+

Outline of Final Research Achievements

We developed a fluorescent probe MGH with a chloroalkyl group, which can be labeled on a HaloTag. By labeling HaloTag expressed in living cells, MGH remained within living cells sufficiently even after 24 h of the treatment. Then, we attempted intracellular Mg2+ imaging during apoptosis, and visualized the free Mg2+ concentration increase after the apoptotic cell shrinkage. Various analyses indicated that the increase in intracellular free Mg2+ concentration during apoptosis was the result of release from Mg-ATP. We also developed fluorescent probes MGQ-1 and MGQ-2 that show high selectivity for Mg2+. The dissociation constants of MGQ-2 for Mg2+ and Ca2+were 0.27 mM and 1.5 mM, respectively, in buffer solution (pH 7.4), and MGQ-2 did not coordinate in the physiological concentration range of Mg-ATP.

Free Research Field

ケミカルバイオロジー