Development of heme oxygenase-1-based heme sensor for quantifying free heme in biological samples

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K01825
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Chemical biology
• Research Institution: Kyushu Institute of Technology
• Principal Investigator: Sakamoto Hiroshi 九州工業大学, 大学院情報工学研究院, 教授 (70309748)
• Co-Investigator(Kenkyū-buntansha): 平 順一 九州工業大学, 大学院情報工学研究院, 助教 (20549612) ; 小松 英幸 九州工業大学, 大学院情報工学研究院, 准教授 (90253567)
• Co-Investigator(Renkei-kenkyūsha): SUEDA Shinji 九州工業大学, 大学院情報工学研究院, 教授 (00325581) ; HIGASHIMOTO Yuichiro 久留米大学, 医学部, 教授 (40352124) ; MORIMATSU Hiroshi 岡山大学, 医学部, 教授 (30379797) ; SHIMIZU Hiroko 岡山大学, 医学部, 客員研究員 (80423284) ; CHIKAZAWA Seishiro 北里大学, 獣医学部, 助教 (80566547) ; SUGAI Manabu 福井大学, 医学部, 教授 (90303891)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: ヘム / センサー / 生体分子 / 酵素 / 計測

Outline of Final Research Achievements

Free heme, the protein-unbound form of heme, participates in a number of physiological events as both a regulatory molecule and a prooxidant species. We previously developed a method for quantifying free heme using fluorescently labeled rat heme oxygenase-1 as a heme sensor. However, quantification of free heme in rat liver microsomal fraction was disturbed by non-specific adhesion of free heme to biomolecules in the sample. To resolve the issue, we investigated the effects of several surfactants on the heme quantification. We also prepared six fluorescent protein-fused rHO-1s and characterized their heme-sensing properties. Among them, three fluorescent protein fusions linked to the N terminus of rHO-1 showed a stoichiometric fluorescent response during titration with heme. Further characterization for intracellular heme detection was performed.

Free Research Field

生物化学