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Temperature dependence of NADPH oxidase in human eosinophils
Author(s) -
Morgan Deri,
Cherny Vladimir V.,
Murphy Ricardo,
Xu Wei,
Thomas Larry L.,
DeCoursey Thomas E.
Publication year - 2003
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2003.041525
Subject(s) - nadph oxidase , superoxide , chemistry , oxidase test , biophysics , arrhenius equation , cytochrome c oxidase , arrhenius plot , enzyme , activation energy , biochemistry , biology
The phagocyte NADPH oxidase helps kill pathogens by producing superoxide anion, O 2 − . This enzyme is electrogenic because it translocates electrons across the membrane, generating an electron current, I e . Using the permeabilized patch voltage‐clamp technique, we studied the temperature dependence of I e in human eosinophils stimulated by phorbol myristate acetate (PMA) from room temperature to > 37°C. For comparison, NADPH oxidase activity was assessed by cytochrome c reduction. The intrinsic temperature dependence of the assembled, functioning NADPH oxidase complex measured during rapid temperature increases to 37°C was surprisingly weak: the Arrhenius activation energy E a was only 14 kcal mol −1 ( Q 10 , 2.2). In contrast, steady‐state NADPH oxidase activity was strongly temperature dependent at 20–30°C, with E a 25.1 kcal mol −1 ( Q 10 , 4.2). The maximum I e measured at 34°C was −30.5 pA. Above 30°C, the temperature dependence of both I e and O 2 − production was less pronounced. Above 37°C, I e was inhibited reversibly. After rapid temperature increases, a secondary increase in I e ensued, suggesting that high temperature promotes assembly of additional NADPH oxidase complexes. Evidently, about twice as many NADPH oxidase complexes are active near 37°C than at 20°C. Thus, the higher Q 10 of steady‐state I e reflects both increased activity of each NADPH oxidase complex and preferential assembly of NADPH oxidase complexes at high temperature. In summary, NADPH oxidase activity in intact human eosinophils is maximal precisely at 37°C.