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. 2010 Oct 18;11(15):2096-9.
doi: 10.1002/cbic.201000115.

Bioluminescent probes of sulfatase activity

Jason S Rush  1 , Kimberly E Beatty, Carolyn R Bertozzi
Affiliations

Bioluminescent probes of sulfatase activity

Jason S Rush et al. Chembiochem. .
No abstract available

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Figures

Figure 1
Figure 1
The caging modules mask luciferin bioluminescence. A. aerogenes sulfatase was incubated with 6.25 μM 1, 2, or 3 for 30 min (sufficient for ~10–20 % aminoluciferin release; see the Supporting Information), after which luminescence was initiated by the addition of luciferase and ATP (gray bars). The addition of H2O instead of sulfatase served as a negative control (white bars). Error bars represent the standard deviation of three replicates. RLU=relative luminescence units.
Figure 2
Figure 2
Sulfatase activity from cell lysates. Gray bars, untreated lysate; White bars, heat-treated lysate; 6.25 μM 1, 2, or 3 was used in all reactions. A) Lysate from E. coli expressing the putative sulfatase YidJ. B) Lysate from M. tuberculosis strain H37Rv. C) Lysate from HEK293 cells at either pH 7.5 or pH 5.6. Luminescence was initiated by the addition of luciferase and ATP. Error bars represent the standard deviation of three replicates.
Figure 3
Figure 3
Sulfatase activity in conditioned media from M. tuberculosis H37Rv and HEK293 cell cultures. Concentrated conditioned medium was incubated with 1 mM 1, 2, or 3 before luminescence was initiated by the addition of luciferase and ATP. For M. tuberculosis, gray bars represent the addition of conditioned medium and white bars represent the addition of fresh, unused medium. For HEK293, gray bars represent conditioned medium from a cell line stably transfected with HSulf2 and white bars represent conditioned medium from parental cells. Error bars represent the standard deviation of three replicates.
Scheme 1
Scheme 1
A) General strategy for the design of sulfatase activated probes. B) Bioluminescent sulfatase reporters.
Scheme 2
Scheme 2
Synthesis of caged aminoluciferin analogues 1–3. a) Cl2CO, dioxane; b) 4, 5, or 6, DMAP, THF; c) D-Cys·HCl, K2CO3, DCM/MeOH/H2O; d) NaN3, DMSO.
See this image and copyright information in PMC

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