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. Author manuscript; available in PMC: 2009 Aug 20.
Published in final edited form as: Bioorg Med Chem Lett. 2008 Sep 10;18(22):5932–5936. doi: 10.1016/j.bmcl.2008.09.021

Figure 4.

Figure 4

‘Click’ chemistry-based ABP labeling of aminopeptidase N. (a) Structure of MH04 probe. (b) Aminopeptidase N labeling was performed as described in Figure 2c using MH04. The ligation of the biotin-azide reporter tag was performed by adding 100 μM of the biotin-azide tag, followed by 1 mM TCEP (tris(2-carboxyethyl) phosphine hydrochloride), 100 μM ligand (tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine) (17× stock in DMSO/tert-butanol 1:4), and 1 mM CuSO4. Reactions were allowed to proceed for 1 h at room temperature, then quenched with equal volume of SDS–PAGE loading buffer. Labeled protein was visualized as in Figure 2b.