Results: 3

1.
Scheme 1

Scheme 1. From: Tags for Labeling Protein N-termini with Subtiligase for Proteomics.

A. Structure of peptide ester 1 with ligation and TEV protease cleavage sites annotated. B. Proteomics workflow for N-terminal labeling using subtiligase.

Hikari A. I. Yoshihara, et al. Bioorg Med Chem Lett. ;18(22):6000-6003.
2.
Figure 2

Figure 2. From: Tags for Labeling Protein N-termini with Subtiligase for Proteomics.

Quantitation of tagging by derivatization and ELISA. A. Tagged proteins are captured on an analytical scale in neutravidin-coated 96-well plates. The propargylglycine residue is derivatized using a copper(I)-catalyzed azide-alkyne cyclization reaction and the incorporated 3-nitrotyrosine is quantitated using an antibody conjugated to horseradish peroxidase (HRP). B. ELISA using peptide ester 3.

Hikari A. I. Yoshihara, et al. Bioorg Med Chem Lett. ;18(22):6000-6003.
3.
Figure 1

Figure 1. From: Tags for Labeling Protein N-termini with Subtiligase for Proteomics.

Avidin blots of ligation reactions in Jurkat cell lysates. A. Comparison of peptide esters at 1 mM and 10 mM. The limited solubility of 1 precludes its use at 10 mM. B. Lysates incubated with increasing concentrations (1, 2, 4, 7 & 10 mM) of peptide ester in the presence of 10 μM subtiligase.

Hikari A. I. Yoshihara, et al. Bioorg Med Chem Lett. ;18(22):6000-6003.

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