title: Monitoring Nitric Oxide in Subcellular Compartments by Hybrid Probe Based on Rhodamine Spirolactam and SNAP-tag.
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.
authors.affiliation: State Key Laboratory of Fine Chemicals and ‡School of Life Science and Technology, Dalian University of Technology , Dalian 116024, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Fine Chemicals and ‡School of Life Science and Technology, Dalian University of Technology , Dalian 116024, China.
authors.forename: Xinbo
authors.initials: X
authors.lastname: Song
authors.affiliation: State Key Laboratory of Fine Chemicals and ‡School of Life Science and Technology, Dalian University of Technology , Dalian 116024, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Han
authors.affiliation: State Key Laboratory of Fine Chemicals and ‡School of Life Science and Technology, Dalian University of Technology , Dalian 116024, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key Laboratory of Fine Chemicals and ‡School of Life Science and Technology, Dalian University of Technology , Dalian 116024, China.
authors.forename: Yongping
authors.initials: Y
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Fine Chemicals and ‡School of Life Science and Technology, Dalian University of Technology , Dalian 116024, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Xiao
mesh_terms: D000818:Animals; D007769:Lactams / Q000737:chemistry*; D051379:Mice; D015335:Molecular Probes / Q000737:chemistry*; D009569:Nitric Oxide / Q000378:metabolism*; D012235:Rhodamines / Q000737:chemistry*; D026423:S-Nitroso-N-Acetylpenicillamine / Q000737:chemistry*; D013347:Subcellular Fractions / Q000378:metabolism*
chemical_list: D007769:Lactams; D015335:Molecular Probes; D012235:Rhodamines; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine