caa a22 4500 475744454 CHVBK 20180406123509.0 cr unu---uuuuu 170329e20001101xx s 000 0 eng 10.1023/A:1007144101361 doi (NATIONALLICENCE)springer-10.1023/A:1007144101361 Conformational Changes in Truncated p47phox Proteins Monitored by Fluorescent Labeling [Elektronische Daten] [Jin Lee, Jeen-Woo Park] The leukocyte NADPH oxidase of neutrophils is a membrane-bound enzyme that catalyzes the reduction of oxygen to O− 2 at the expense of NADPH. During activation, the cytosolic oxidase components p47phox and p67phox, each containing two Src homology 3 (SH3) domains, migrate to the plasma membrane. p47phox and p67phox associate with cytochrome b558, a membrane-integrated flavohemoprotein, to assemble the active oxidase. Oxidase activation can be mimicked in a cell-free system using an anionic amphiphile, such as sodium dodecyl sulfate or arachidonic acid, as an activating agent. Activators of the oxidase in vitro cause exposure of the SH3 domains of p47phox, which has probably been masked by the C-terminal region of this protein in a resting state. We show here that the fluorescence exhibited by the covalently labeled N,N′-di-methyl-N(iodoacetyl)-N′-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) ethyleneamine (IANBD) was increased when N-terminal-truncated p47phox-(SH3)2-C was treated with anionic amphiphiles. This finding was similar to the results obtained with the full-length p47phox. However, the fluorescence of C-terminal-truncated p47phox-N-(SH3)2 and that of both C-terminal and N-terminal truncated p47phox-(SH3)2 were not altered by the activators. These results indicate that the C-terminal region of p47phox is a primary target of the conformational change during the activation of NADPH oxidase. Plenum Publishing Corporation, 2000 Truncated p47phox proteins nationallicence SH3 domains nationallicence fluorescent labeling nationallicence amphiphile nationallicence Lee Jin Department of Biochemistry, College of Natural Sciences, Kyungpook National University, 702-701, Taegu, Korea aut Park Jeen-Woo Department of Biochemistry, College of Natural Sciences, Kyungpook National University, 702-701, Taegu, Korea aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/8(2000-11-01), 643-648 0277-8033 19:8<643 2000 19 10930 https://doi.org/10.1023/A:1007144101361 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007144101361 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lee Jin Department of Biochemistry, College of Natural Sciences, Kyungpook National University, 702-701, Taegu, Korea aut NATIONALLICENCE 700 1- Park Jeen-Woo Department of Biochemistry, College of Natural Sciences, Kyungpook National University, 702-701, Taegu, Korea aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/8(2000-11-01), 643-648 0277-8033 19:8<643 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer