title: Effects of pO2 on the activation of skeletal muscle ryanodine receptors by NO: a cautionary note.
abstract: Eu et al., reported that O2 dynamically controls the redox state of 6-8 out of 50 thiols per skeletal ryanodine receptor (RyR1) subunit and thereby tunes the response of Ca2+-release channels to authentic nitric oxide (NO) [J.P. Eu, J. Sun, L. Xu, J.S. Stamler, G. Meissner, The skeletal muscle calcium release channel: coupled O2 sensor and NO signaling functions, Cell 102 (2000) 499-509]. A role for O2 was based on the observation that RyR1 can be activated by submicromolar NO at physiological ( approximately 10 mmHg) but not ambient (approximately 150 mmHg) pO2. At ambient pO2, these critical thiols were oxidized but incubation at low pO2 reset the redox state of these thiols, closed RyR1 channels and made these thiols available for nitrosation by low NO concentrations. Eu et al., postulated the existence of a redox/O2sensor that couples channel activity to NO and pO2 and explained that "the nature of the 'redox/O2 sensor' that couples channel activity to intracellular redox chemistry is a mystery". Here, we re-examined the effect of pO2 on RyR1 and find that incubation of RyR1 at low pO2 did not alter channel activity and NO (0.5-50 microM) failed to activate RyR1 despite a wide range of pO2 pre-incubation conditions. We show that low levels of NO do not activate RyR1, do not reverse the inhibition of RyR1 by calmodulin (CaM) even at physiological pO2. Similarly, the pre-incubation of SR vesicles in low pO2 (for 10-80 min) did not inhibit channel activity or sensitization of RyR1 to NO. We discuss the significance of these findings and propose that caution should be taken when considering a role for pO2 and nitrosation by NO as mechanisms that tune RyRs in striated muscles.
authors.affiliation: Department of Cell Biology and Physiology, University of Pittsburgh, School of Medicine, 3500 Terrace Street, Room S314 Biomedical Science Tower, Pittsburgh, PA 15261, USA.
authors.forename: Eunji
authors.initials: E
authors.lastname: Cheong
authors.forename: Vassil
authors.initials: V
authors.lastname: Tumbev
authors.forename: Detcho
authors.initials: D
authors.lastname: Stoyanovsky
authors.forename: Guy
authors.initials: G
authors.lastname: Salama
mesh_terms: D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism*; D009569:Nitric Oxide / Q000494:pharmacology*; D010100:Oxygen / Q000378:metabolism / Q000494:pharmacology*; D010313:Partial Pressure; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects / Q000378:metabolism*
chemical_list: D019837:Ryanodine Receptor Calcium Release Channel; D009569:Nitric Oxide; D010100:Oxygen