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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

Supelco

2-Butanone, analytical standard

Sigma-Aldrich

2-Butanone, ACS reagent, ≥99.0%

Sigma-Aldrich

2-Butanone, ReagentPlus^®, ≥99%

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2-Butanone, suitable for HPLC, ≥99.7%

Sigma-Aldrich

2-Butanone, ACS reagent, ≥99.0%

Millipore

Bifido Selective Supplement B, suitable for microbiology

USP

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

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5α-Androstan-17β-ol-3-one, ≥97.5%

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

Supelco

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

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

Supelco

Acetic acid, analytical standard

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Acetic acid, for luminescence, BioUltra, ≥99.5% (GC)

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

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

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Magnesium sulfate, ≥99.99% trace metals basis

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

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Tyramine, ≥98.0%

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2-Butanone, FCC, FG

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

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

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Magnesium sulfate solution, BioUltra, for molecular biology

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Acetyl chloride, reagent grade, 98%

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Acetyl chloride, reagent grade, 98%

Supelco

Methyl Ethyl Ketone, Pharmaceutical Secondary Standard; Certified Reference Material

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

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Acetyl chloride solution, 1 M in methylene chloride

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

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