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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

2-Butanone, suitable for HPLC, ≥99.7%

Sigma-Aldrich

Lithium bromide, ReagentPlus^®, ≥99%

Sigma-Aldrich

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

Sigma-Aldrich

Magnesium sulfate solution, BioUltra

Sigma-Aldrich

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

Sigma-Aldrich

Acetyl chloride, reagent grade, 98%

Sigma-Aldrich

Tyramine, 98%, FG

Millipore

Bifido Selective Supplement B, suitable for microbiology

Supelco

Tyramine, analytical standard

Sigma-Aldrich

2-Butanone, ACS reagent, ≥99.0%

Sigma-Aldrich

2-Butanone, ACS reagent, ≥99.0%

Supelco

Methyl Ethyl Ketone, Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

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

Sigma-Aldrich

2-Butanone, ReagentPlus^®, ≥99%

Sigma-Aldrich

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

Supelco

2-Butanone, analytical standard

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

Magnesium sulfate solution, 1.00 M±0.04 M

Sigma-Aldrich

5α-Androstan-17β-ol-3-one, ≥97.5%

Sigma-Aldrich

Acetyl chloride solution, 1 M in methylene chloride

Sigma-Aldrich

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

Sigma-Aldrich

2-Butanone, JIS special grade, ≥99.0%

Sigma-Aldrich

2-Butanone, SAJ first grade, ≥99.0%

Sigma-Aldrich

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

Sigma-Aldrich

2-Butanone, FCC, FG

Supelco

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

Sigma-Aldrich

Tyramine, ≥98.0%

Sigma-Aldrich

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

USP

Glacial acetic acid, United States Pharmacopeia (USP) Reference Standard