Nbd glycerol 3 pea

The enzymatic assay was performed according to a previous reported method with a slight modification^{38 (link)}. Briefly, 0.1 mM full-length MCR-1 was treated with 10 molar equivalents of AuCl or Au(PEt₃)Cl at 25 °C for 18 h and subsequently incubated with 0.1 mM of a fluorescent substrate, nitrobenzodiazole-labeled glycerol-3-phosphoethanolamine (NBD-glycerol-3-PEA, Avanti Lipids, USA) in assay buffer (50 mM HEPES, pH 7.5, 100 mM NaAc, 0.023% DDM) at 25 °C overnight. TLC plate was used to separate NBD-glycerol from the MCR-1 reaction mixture in a mobile phase [ethyl acetate: methanol: water, 7:2:1 (vol/vol)]. The reaction product was analyzed by exposing the TLC plate to UV light (455-485 nm) and visualizing the fluorescent signals with a gel imaging system.

Meropenem was purchased from TCI Chemicals (Shanghai, China). Kanamycin sulfate and Luria–Bertani (LB) broth powder were purchased from Affymetrix. Auranofin (AUR) was purchased from MedChemExpress (MCE, USA). The fluorescent substrate NBD-glycerol-3-PEA was purchased from Avanti Lipids, USA. TLC plate (Silica gel 60 F₂₅₄, Aluminium sheets, 20 × 20 cm) was purchased from Merck, Germany. All the other chemicals were purchased from Sigma-Aldrich unless otherwise stated. All the sources of bacteria employed for cell-based and animal studies are listed in Supplementary Table 7. All the specific primers for different genes listed in Supplementary Table 8 were synthesized by BGI Gene Company (Shenzhen, China).