TY - JOUR
T1 - A new role of Pro-73 of p47phox in the activation of neutrophil NADPH oxidase
AU - Nagasawa, Teruaki
AU - Ebisu, Kentaro
AU - Inoue, Yasuyuki
AU - Miyano, Kei
AU - Tamura, Minoru
N1 - Funding Information:
This work was supported in part by a grant (13480297) for scientific research from the Ministry of Education, Science, Sports and Culture of Japan. We are grateful to Shuhei Kobayashi, Mamoru Tanaka, Yasuaki Komeima, Tetsutaro Kai, and Kyoji Watanabe for their excellent technical assistance and to Drs. Koichi Tanaka, Yaeta Endo, and Keiichi Kato (Department of Applied Chemistry, Ehime University) for their discussions and instruments. We are grateful to Drs. Chang-Hoon Han (Department of Biochemistry, ETH-Zentrum), Dave Lambeth (Department of Pathology, Emory University), and Toshitsugu Yubisui (Department of Biochemistry, Okayama University of Science) for their cDNA clones and helpful discussions.
PY - 2003/8/1
Y1 - 2003/8/1
N2 - The PX domain of p47phox is thought to be involved in autoinhibition. However, when the domain was deleted, the ability to activate the phagocyte NADPH oxidase was markedly diminished. We have mutated the proline-rich region of the PX domain and examined the mutants for the ability to activate. Substitution of Gln for Pro-73 of p47phox(1-286) (P73Q) resulted in a considerably lower activity than the wild type and P73Q had a much lower affinity for the oxidase complex. Whereas, Gln substitution for Pro-76 (P76Q) showed a slightly enhanced activation and the mutant had a slightly higher affinity for the complex than the wild type. Affinity for p67phox(1-210) was slightly decreased either by P73Q or P76Q. Optimal SDS concentration for the activation was lowered by these mutations. Binding of PX domain with phosphatidylinositol-3,4-bisphosphate was diminished by P73Q mutation. The results in this study suggest that Pro-73 has a role in interaction with the catalytic component cytochrome b558.
AB - The PX domain of p47phox is thought to be involved in autoinhibition. However, when the domain was deleted, the ability to activate the phagocyte NADPH oxidase was markedly diminished. We have mutated the proline-rich region of the PX domain and examined the mutants for the ability to activate. Substitution of Gln for Pro-73 of p47phox(1-286) (P73Q) resulted in a considerably lower activity than the wild type and P73Q had a much lower affinity for the oxidase complex. Whereas, Gln substitution for Pro-76 (P76Q) showed a slightly enhanced activation and the mutant had a slightly higher affinity for the complex than the wild type. Affinity for p67phox(1-210) was slightly decreased either by P73Q or P76Q. Optimal SDS concentration for the activation was lowered by these mutations. Binding of PX domain with phosphatidylinositol-3,4-bisphosphate was diminished by P73Q mutation. The results in this study suggest that Pro-73 has a role in interaction with the catalytic component cytochrome b558.
UR - http://www.scopus.com/inward/record.url?scp=0037676127&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037676127&partnerID=8YFLogxK
U2 - 10.1016/S0003-9861(03)00296-0
DO - 10.1016/S0003-9861(03)00296-0
M3 - Article
C2 - 12859985
AN - SCOPUS:0037676127
SN - 0003-9861
VL - 416
SP - 92
EP - 100
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 1
ER -