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Monitoring Nitric Oxide in Subcellular Compartments by Hybrid Probe Based on Rhodamine Spirolactam and SNAP-tag
- Source :
- ACS Chemical Biology. 11:2033-2040
- Publication Year :
- 2016
- Publisher :
- American Chemical Society (ACS), 2016.
-
Abstract
- By connection of O(6)-benzylguanine (BG) to an "o-phenylenediamine-locked" rhodamine spirolactam responsive to nitric oxide (NO), a novel substrate (TMR-NO-BG) of genetically encoded SNAP-tag has been constructed. In living cells, labeling SNAP-tag fused proteins with TMR-NO-BG will in situ generate corresponding probe-protein conjugates (TMR-NO-SNAP) that not only inherit high NO sensitivity from the small-molecule parent but also guarantee the site-specificity to the designated subcellular compartments such as the mitochondrial inner membrane, nucleus, and cytoplasm. In two representative cellular processes, TMR-NO-BG demonstrates its applicability to monitor endogenous subcellular NO in the activated RAW264.7 cells stimulated by lipopolysaccharide and in the apoptotic COS-7 cells induced by etoposide.
- Subjects :
- In situ
Lactams
S-Nitroso-N-Acetylpenicillamine
Biology
Nitric Oxide
010402 general chemistry
01 natural sciences
Biochemistry
Nitric oxide
Rhodamine
Mice
chemistry.chemical_compound
Animals
Inner mitochondrial membrane
Rhodamines
010405 organic chemistry
General Medicine
0104 chemical sciences
SNAP-tag
chemistry
Cytoplasm
Molecular Probes
Biophysics
Molecular Medicine
S-Nitroso-N-acetylpenicillamine
Molecular probe
Subcellular Fractions
Subjects
Details
- ISSN :
- 15548937 and 15548929
- Volume :
- 11
- Database :
- OpenAIRE
- Journal :
- ACS Chemical Biology
- Accession number :
- edsair.doi.dedup.....1a863d34b310a7897a0c6aa78956f58b