Fluorescently Modified NDM-1: A Versatile Drug Sensor for Rapid β-Lactam Antibiotic and Inhibitor Screening

Overview

Journal ACS Omega

Publisher American Chemical Society

Specialty Chemistry

Date 2024 Mar 4

PMID 38434906

Authors

Sai-Fung Chung

Suet-Ying Tam

Wai-Ting Wong

Pui-Kin So

Wing-Lam Cheong

Chun-Wing Mak

Leo Man-Yuen Lee

Pak-Ho Chan

Kwok-Yin Wong

Yun-Chung Leung

Affiliations

Soon will be listed here.

Abstract

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 drug screening.

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