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. 2005 Oct 4;2(10):e295. doi: 10.1371/journal.pmed.0020295

Figure 1. P. pastoris Secrete Ac-APR-1 Zymogen that Autoactivates at Low pH and Degrades Canine Hb.

Figure 1

SDS-PAGE gel stained with Coomassie Brilliant Blue showing purification of recombinant APR-1 zymogen from P. pastoris culture supernatant.

(A) Lane 1, molecular weight markers; lane 2, concentrated culture supernatant; lane 3, flow-through from a nickel-IDA column; lane 4, 5 mM imidazole wash; lane 5, 20 mM imidazole column eluate; lane 6, 60 mM imidazole eluate; and lane 7, 1 M imidazole eluate. Purified recombinant APR-1 zymogen was activated by buffer exchange into 0.1 M sodium formate/0.1 M NaCl (pH 3.6).

(B) Lane 1, molecular weight markers; lane 2, 5.0 μg of canine Hb (pH 3.6); and lane 3, 5.0 μg of canine Hb (pH 3.6) incubated with 0.2 μg of recombinant APR-1.