Improvement of heme oxygenase-1-based heme sensor for quantifying free heme in biological samples
Improvement of heme oxygenase-1-based heme sensor for quantifying free heme in biological samples
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DOI：
10.1016/j.ab.2015.08.004
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发表时间：
2015-11-15
影响因子：
2.9
通讯作者：
Sakamoto, Hiroshi
中科院分区：
文献类型：
--
作者：
Taira, Junichi;Nakashima, Yukinori;Sakamoto, Hiroshi
我们最近报道了一种使用荧光标记血红素氧化酶-1的新型血红素传感器；然而，其固有的酶活性将成为定量生物样品中血红素的潜在障碍。在这里，我们发现具有重要催化作用的残基Asp140与传感器中的组氨酸发生突变，不仅降低了血红素降解活性，而且增加了血红素结合亲和力。具有可见荧光团的传感器还有助于避免生物样品中内源性物质的本底发射。利用改进的血红素传感器，我们首次成功地定量测定了大鼠肝脏样品中的游离血红素。(C) 2015爱思唯尔公司版权所有。
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. (C) 2015 Elsevier Inc. All rights reserved.