TY - JOUR
T1 - Post-translational tyrosine nitration of eosinophil granule toxins mediated by eosinophil peroxidase
AU - Ulrich, Martina
AU - Petre, Alina
AU - Youhnovski, Nikolay
AU - Prömm, Franziska
AU - Schirle, Markus
AU - Schumm, Michael
AU - Pero, Ralph S.
AU - Doyle, Alfred
AU - Checkel, James
AU - Kita, Hirohito
AU - Thiyagarajan, Nethaji
AU - Acharya, K. Ravi
AU - Schmid-Grendelmeier, Peter
AU - Simon, Hans Uwe
AU - Schwarz, Heinz
AU - Tsutsui, Masato
AU - Shimokawa, Hiroaki
AU - Bellon, Gabriel
AU - Lee, James J.
AU - Przybylski, Michael
AU - Döring, Gerd
PY - 2008/10/17
Y1 - 2008/10/17
N2 - Nitration of tyrosine residues has been observed during various acute and chronic inflammatory diseases. However, the mechanism of tyrosine nitration and the nature of the proteins that become tyrosine nitrated during inflammation remain unclear. Here we show that eosinophils but not other cell types including neutrophils contain nitrotyrosine-positive proteins in specific granules. Furthermore, we demonstrate that the human eosinophil toxins, eosinophil peroxidase (EPO), major basic protein, eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP), and the respective murine toxins, are post-translationally modified by nitration at tyrosine residues during cell maturation. High resolution affinity-mass spectrometry identified specific single nitration sites at Tyr349 in EPO and Tyr33 in both ECP and EDN. ECP and EDN crystal structures revealed and EPO structure modeling suggested that the nitrated tyrosine residues in the toxins are surface exposed. Studies in EPO-/-, gp91phox-/-, and NOS-/- mice revealed that tyrosine nitration of these toxins is mediated by EPO in the presence of hydrogen peroxide and minute amounts of NOx. Tyrosine nitration of eosinophil granule toxins occurs during maturation of eosinophils, independent of inflammation. These results provide evidence that post-translational tyrosine nitration is unique to eosinophils.
AB - Nitration of tyrosine residues has been observed during various acute and chronic inflammatory diseases. However, the mechanism of tyrosine nitration and the nature of the proteins that become tyrosine nitrated during inflammation remain unclear. Here we show that eosinophils but not other cell types including neutrophils contain nitrotyrosine-positive proteins in specific granules. Furthermore, we demonstrate that the human eosinophil toxins, eosinophil peroxidase (EPO), major basic protein, eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP), and the respective murine toxins, are post-translationally modified by nitration at tyrosine residues during cell maturation. High resolution affinity-mass spectrometry identified specific single nitration sites at Tyr349 in EPO and Tyr33 in both ECP and EDN. ECP and EDN crystal structures revealed and EPO structure modeling suggested that the nitrated tyrosine residues in the toxins are surface exposed. Studies in EPO-/-, gp91phox-/-, and NOS-/- mice revealed that tyrosine nitration of these toxins is mediated by EPO in the presence of hydrogen peroxide and minute amounts of NOx. Tyrosine nitration of eosinophil granule toxins occurs during maturation of eosinophils, independent of inflammation. These results provide evidence that post-translational tyrosine nitration is unique to eosinophils.
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U2 - 10.1074/jbc.M801196200
DO - 10.1074/jbc.M801196200
M3 - Article
C2 - 18694936
AN - SCOPUS:57649120776
SN - 0021-9258
VL - 283
SP - 28629
EP - 28640
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 42
ER -