抗生素
体外
内酰胺
化学
药品
药理学
微生物学
组合化学
医学
立体化学
生物化学
生物
作者
Sai-Fung Chung,Suet-Ying Tam,Wai-Ting Wong,Pui‐Kin So,Wing‐Lam Cheong,Chee Leung Mak,Leo Man-Yuen Lee,Pak‐Ho Chan,Kwok Yin Wong,Yun-Chung Leung
出处
期刊:ACS omega
[American Chemical Society]
日期:2024-02-13
标识
DOI:10.1021/acsomega.3c08117
摘要
We successfully developed a fluorescent drug sensor from clinically relevant New Delhi metallo-β-lactamase-1 (NDM-1). The F70 residue was chosen to be replaced with a cysteine for conjugation with thiol-reactive fluorescein-5-maleimide to form fluorescent F70Cf, where “f” refers to fluorescein-5-maleimide. Our proteolytic studies of unlabeled F70C and labeled F70Cf monitored by electrospray ionization–mass spectrometry (ESI-MS) revealed that fluorescein-5-maleimide was specifically linked to C70 in 1:1 mole ratio (F70C:fluorophore). Our drug sensor (F70Cf) can detect the β-lactam antibiotics cefotaxime and cephalothin by giving stronger fluorescence in the initial binding phase and then declining fluorescence signals as a result of the hydrolysis of the antibiotics into acid products. F70Cf can also detect non-β-lactam inhibitors (e.g., l-captopril, d-captopril, dl-thiorphan, and thanatin). In all cases, F70Cf exhibits stronger fluorescence due to inhibitor binding and subsequently sustained fluorescence signals in a later stage. Native ESI-MS results show that F70Cf can bind to all four inhibitors. Moreover, our drug sensor is compatible with a high-throughput microplate reader and has the capability to perform in vitro drug screening.
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