Figure 4.

MINDY1 uses a non-canonical catalytic mechanism

(A) Close-up view of the catalytic site in MINDY1 apo. The dotted red lines indicate hydrogen bonds, the dotted black line the ionic bond, and the blue sphere indicates water molecule.

(B) DUB assay comparing the cleavage of fluorescently labeled K48-Ub₅ by MINDY1 WT, S321A, and S321D mutants. The percentage of pentaUb hydrolyzed is plotted against time (right). n = 2; mean ± SD.

(C) Close-up view of the catalytic site in MINDY1^C137A:K48-Ub₂ complex.

(D) DUB assay as in (B) comparing the chain cleavage by MINDY1 WT, T335V, and T335D mutants. The percentage of pentaUb hydrolyzed is plotted against time (right). n = 2; mean ± SD.

(E) Close-up view of the catalytic site in MINDY1^T335D.

See also Figure S5.