Skip to main content
. 2015 Mar 20;290(19):12135–12146. doi: 10.1074/jbc.M114.624072

FIGURE 3.

FIGURE 3.

Purification and circular dichroism of recombinant ADAM17 proenzyme WT and mutants. A, representative SDS-PAGE of purification of ADAM17 proenzyme with a nickel-nitrilotriacetic acid column followed by anion exchange column: molecular mass (MW), nickel-nitrilotriacetic acid lysate load (1), nickel-nitrilotriacetic acid flow-through (2), anion exchange load (3), anion exchange flow-through (4), and anion exchange first peak (5). B, schematic presentation of ADAM17 domain structure; SP, signal peptide; PRO, pro-domain; CAT, catalytic domain; DIS, disintegrin domain; CYT, cytoplasmic tail. The blue arrow indicates the upstream PC site and the red arrow indicates the boundary site. Below are the recombinant ADAM17 proenzyme constructs with an N-terminal His tag: WT, pro-catalytic domains; US-R58A mutation in the Upstream PC site; BS, R211A/R214G mutation in the boundary site; and 2M, R58A/R211A/R214G containing both PC sites mutations. C, CD measurements of WT ADAM17 proenzyme and mutants.