Improvement of heme oxygenase-1-based heme sensor for quantifying free heme in biological samples |
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| Authors: | Junichi Taira Yukinori Nakashima Shun Yoshihara Shinya Koga Shinji Sueda Hideyuki Komatsu Yuichiro Higashimoto Toru Takahashi Nohito Tanioka Hiroko Shimizu Hiroshi Morimatsu Hiroshi Sakamoto |
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| Affiliation: | 1. Department of Bioscience and Bioinformatics, Graduate School of Computer Science and Systems Engineering, Kyushu Institute of Technology, Kawazu, Iizuka 820-8502, Japan;2. Department of Chemistry, Kurume University School of Medicine, Asahi-machi, Kurume 830-0011, Japan;3. Faculty of Health and Welfare Science, Okayama Prefectural University, Kuboki, Okayama 719-1197, Japan;4. Department of Anesthesiology and Resuscitology, Okayama University Medical School, Shikatacho, Okayama 700-8558, Japan |
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| Abstract: | We recently reported a novel heme sensor using fluorescently labeled heme oxygenase-1; however, its inherent enzyme activity would be a potential obstacle in quantifying heme in biological samples. Here, we found that mutation of the catalytically important residue, Asp140, with histidine in the sensor not only diminished the heme degradation activity but also increased heme binding affinity. The sensor with a visible fluorophore was also found to be beneficial to avoid background emission from endogenous substance in biological samples. By using the improved heme sensor, we succeeded in quantifying free heme in rat hepatic samples for the first time. |
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| Keywords: | Biological sample Fluorescent labeling Free heme Heme degradation Heme oxygenase-1 |
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