| Autor: |
Tallman KR; Program in Chemical Biology and the Department of Biomedical Engineering, Oregon Health & Science University , Portland, Oregon 97201, United States., Levine SR; Program in Chemical Biology and the Department of Biomedical Engineering, Oregon Health & Science University , Portland, Oregon 97201, United States., Beatty KE; Program in Chemical Biology and the Department of Biomedical Engineering, Oregon Health & Science University , Portland, Oregon 97201, United States. |
| Jazyk: |
angličtina |
| Zdroj: |
ACS chemical biology [ACS Chem Biol] 2016 Jul 15; Vol. 11 (7), pp. 1810-5. Date of Electronic Publication: 2016 May 23. |
| DOI: |
10.1021/acschembio.6b00233 |
| Abstrakt: |
Enzyme-activated, fluorogenic probes are powerful tools for studying bacterial pathogens, including Mycobacterium tuberculosis (Mtb). In prior work, we reported two 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one) (DDAO)-derived acetoxymethyl ether probes for esterase and lipase detection. Here, we report four-carbon (C4) and eight-carbon (C8) acyloxymethyl ether derivatives, which are longer-chain fluorogenic substrates. These new probes demonstrate greater stability and lipase reactivity than the two-carbon (C2) acetoxymethyl ether-masked substrates. We used these new C4 and C8 probes to profile esterases and lipases from Mtb. The C8-masked probes revealed a new esterase band in gel-resolved Mtb lysates that was not present in lysates from nonpathogenic M. bovis (bacillus Calmette-Guérin), a close genetic relative. We identified this Mtb-specific enzyme as the secreted esterase Culp1 (Rv1984c). Our C4- and C8-masked probes also produced distinct Mtb banding patterns in lysates from Mtb-infected macrophages, demonstrating the potential of these probes for detecting Mtb esterases that are active during infections. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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