DNA from bronchial secretions modulates elastase inhibition by alpha(1)-proteinase inhibitor and oxidized secretory leukoprotease inhibitor.

Abstract

Previously we reported that DNA from sputum promotes the inhibition of human leukocyte elastase (HLE) by native secretory leukoprotease inhibitor (SLPI). This study shows that sputum DNA also promotes the inhibition by oxidized SLPI, a form of SLPI that may occupy a large fraction of the inhibitor in the lungs under conditions of high oxidative stress. With sputum DNA at 5 microg/ml, a concentration much lower than those in vivo, the inhibition constant (K(i) ) of oxidized SLPI against HLE is reduced from 31 nM to 23 to 920 pM, as compared with the K(i) of native SLPI, 58 pM, under the same conditions. On the other hand, sputum DNA retards inhibition of HLE by alpha(1)-proteinase inhibitor (alpha(1)-PI). The association rate of alpha(1)-PI and HLE is decreased from 1 x 10(7) M(-1) s(-1) in the absence of DNA to 2 to 6 x 10(6) M(-1) s(-1) in the presence of sputum DNA at 100 microg/ml. On the basis of results with an elastase-specific oligonucleotide aptamer, it was found that the downregulation of alpha(1)-PI activity can be attributed to an interaction between sputum DNA and multiple DNA-binding sites on HLE. DNA-binding sites on HLE also participate in the upregulation of oxidized SLPI activity. Data from this and our previous studies demonstrate that sputum DNA facilitates the association of HLE with native and oxidized SLPI, whereas it delays the association of HLE with alpha(1)-PI. We conclude that by modulating the inhibition of HLE, sputum DNA directly affects the balance between proteases and antiproteases in the lungs.