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Mixed alkyl aryl phosphonate esters as quenched fluorescent activity-based probes for serine proteases.

Mixed alkyl aryl phosphonate esters as quenched fluorescent activity-based probes for serine proteases.

Organic & biomolecular chemistry (2015-01-03)

Sevnur Serim, Philipp Baer, Steven H L Verhelst

ABSTRACT

Activity-based probes (ABPs) are powerful tools for the analysis of active enzyme species in whole proteomes, cells or animals. Quenched fluorescent ABPs (qABPs) can be applied for real time imaging, allowing the visualization of dynamic enzyme activation by fluorescent microscopy. Unfortunately, qABPs are only available for a few enzymes. We here describe the design and synthesis of qABPs for serine proteases based on a phosphonate ester scaffold.

MATERIALS

Product Number

Brand

Product Description

Sigma-Aldrich

Acetyl chloride solution, 1 M in methylene chloride

Sigma-Aldrich

Di-tert-butyl dicarbonate solution, 2 M in THF

Sigma-Aldrich

Tyramine, 98%, FG

Millipore

Bifido Selective Supplement B, suitable for microbiology

Supelco

5α-Androstan-17β-ol-3-one, VETRANAL^®, analytical standard

Sigma-Aldrich

Di-tert-butyl dicarbonate solution, 2 M in methylene chloride

Sigma-Aldrich

Lithium bromide, AnhydroBeads^™, −10 mesh, 99.999% trace metals basis

Supelco

2-Butanone, analytical standard

Sigma-Aldrich

5α-Androstan-17β-ol-3-one, purum, ≥99.0% (TLC)

Sigma-Aldrich

Magnesium sulfate, ≥99.99% trace metals basis

Sigma-Aldrich

Lithium bromide, AnhydroBeads^™, −10 mesh, ≥99.9% trace metals basis

Sigma-Aldrich

Di-tert-butyl dicarbonate solution, 1.0 M in THF

Sigma-Aldrich

2-Butanone, FCC, FG

Sigma-Aldrich

Lithium bromide, powder and chunks, ≥99.995% trace metals basis

Sigma-Aldrich

Magnesium sulfate solution, BioUltra, for molecular biology

Sigma-Aldrich

Tyramine, ≥98.0%

Sigma-Aldrich

Magnesium sulfate, BioReagent, suitable for cell culture, suitable for insect cell culture

Sigma-Aldrich

5α-Androstan-17β-ol-3-one, ≥97.5%

Sigma-Aldrich

Magnesium sulfate solution, for molecular biology, 1.00 M±0.04 M

Sigma-Aldrich

Acetyl chloride, reagent grade, 98%

Sigma-Aldrich

Acetyl chloride, puriss. p.a., ≥99.0% (T)

Sigma-Aldrich

Acetyl chloride, reagent grade, 98%

Sigma-Aldrich

2-Butanone, ACS reagent, ≥99.0%

Sigma-Aldrich

Magnesium sulfate, puriss. p.a., drying agent, anhydrous, ≥98.0% (KT), powder (very fine)

Sigma-Aldrich

Lithium bromide, anhydrous, free-flowing, Redi-Dri^™, ReagentPlus^®, ≥99%

Sigma-Aldrich

Magnesium sulfate, anhydrous, free-flowing, Redi-Dri^™, ReagentPlus^®, ≥99.5%

Sigma-Aldrich

Magnesium sulfate, anhydrous, ReagentPlus^®, ≥99.5%

Supelco

Tyramine, analytical standard

Sigma-Aldrich

Magnesium sulfate, anhydrous, reagent grade, ≥97%

Sigma-Aldrich

2-Butanone, ReagentPlus^®, ≥99%