Figure 4: Impaired inhibition of tc mouse neutrophil elastase by AAT. | Nature Communications

Figure 4: Impaired inhibition of tc mouse neutrophil elastase by AAT.

From: Autoprocessing of neutrophil elastase near its active site reduces the efficiency of natural and synthetic elastase inhibitors

Figure 4

Wild-type or mutated mouse neutrophil elastase (wt mNE or mut mNE) was mixed with AAT at a ratio of 2:3 and incubated at 37 °C. A covalent bond is formed between the Ser195 of neutrophil elastase and the Met358 of AAT. To study the complex formation over time, aliquots were withdrawn at the indicated time points and analysed by SDS–PAGE and Coomassie staining. (a) The tc neutrophil elastase–AAT complex (tc-AAT) is smaller than the sc neutrophil elastase–AAT complex (sc-AAT) as only the small 7-kDa carboxyl-terminal fragment of tc mouse neutrophil elastase remains covalently bound to AAT in reducing conditions. While sc neutrophil elastase–AAT complexes form within seconds, tc neutrophil elastase–AAT complex formation takes several minutes. (b) Mutated mouse neutrophil elastase consists of almost exclusively sc neutrophil elastase. The reaction with AAT is finished after ten seconds. Both experiments were repeated thrice.

Back to article page