Z vlr amc

Proteasome substrates: The fluorogenic substrates Suc-LLVY-AMC, Z-LLE-AMC, Z-VLR-AMC, and Ac-ANW-AMC were from Boston Biochem (Cambridge, MA). Human immunoproteasomes (isolated from peripheral blood mononuclear cells) and human constitutive proteasomes (isolated from red blood cells) were from Boston Biochem Inc. Mtb20SOG was expressed and purified as reported ^{10 (link)}.

Suc-LLVY-AMC, Ac-ANW-AMC, Z-VLR-AMC, Ac-LLE-AMC, Ac-PAL-AMC, human c-20S (RBC) and i-20S (PBMC) were from Boston Biochem (Cambridge, MA). Enzymatic assays were recorded on a Molecular Devices SpectraMax M5 plate readers. The human blood samples were sourced ethically and their research use was in accord with the terms of the informed consent. Purity of all final compounds were > 95%.

The fluorogenic substrate Suc-LLVY-AMC, Z-LLE-AMC, and Z-VLR-AMC were from Boston Biochem (Cambridge, MA). Human immunoproteasomes (i-20S, isolated from peripheral blood mononuclear cells), human constitutive proteasomes (c-20S, isolated from red blood cells), and recombinant human PA28 activator alpha subunit (PA28α) were from Boston Biochem Inc. Mtb20SOG was expressed and purified as reported.^{27 (link)} Mouse and human liver microsomes were purchased from Sekisui XenoTech, LLC. (Kansas City, KS). Purity of the compounds was determined on a Waters Acquity^™ UPLC / MS system equipped with a C18 column (100 × 2.1 mm, 1.7 μm), coupled with a PDA detector. The purity of all final compounds was > 95% except macrocycle 13. The purity of macrocycle 13 was 93%.