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. Author manuscript; available in PMC: 2012 Jun 4.
Published in final edited form as: J Biol Chem. 2007 Jun 1;282(32):23129–23139. doi: 10.1074/jbc.M701857200

FIGURE 6. An olefin intermediate modifies the active site thiol following sortase activation of AAEK1.

FIGURE 6

A, proposed structure of the olefin intermediate and its reaction with thiol. B, reactions were performed in the presence or absence of DTT and the degree of peptide cleavage assayed 1 h later, as described for supplemental Fig. S1. The symbols (+ and −) represent the presence or absence of the indicated reagents, whereas the numbers refer to the concentration of DTT. The far right reaction “1hr” refers to the addition of DTT to the sample 1 h after the reaction with inhibitor was started. C, reactions were performed as in B, except that the concentration of AAEK1 and the putative elimination intermediates 11 and 12 were assayed at 5 and 50 μM. The mean ±S.D. of three independent experiments are presented in B and C. Ar, thiophen-2-yl.