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

Junichi Taira, Yukinori Nakashima, Shun Yoshihara, Shinya Koga, Shinji Sueda, Hideyuki Komatsu, Yuichiro Higashimoto, Toru Takahashi, Nohito Tanioka, Hiroko Shimizu, Hiroshi Morimatsu, Hiroshi Sakamoto

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)50-52
Number of pages3
JournalAnalytical Biochemistry
Volume489
DOIs
Publication statusPublished - Nov 15 2015

Keywords

  • Biological sample
  • Fluorescent labeling
  • Free heme
  • Heme degradation
  • Heme oxygenase-1

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Taira, J., Nakashima, Y., Yoshihara, S., Koga, S., Sueda, S., Komatsu, H., Higashimoto, Y., Takahashi, T., Tanioka, N., Shimizu, H., Morimatsu, H., & Sakamoto, H. (2015). Improvement of heme oxygenase-1-based heme sensor for quantifying free heme in biological samples. Analytical Biochemistry, 489, 50-52. https://doi.org/10.1016/j.ab.2015.08.004