© 1995 American Heart Association, Inc.
Articles |
Functional Consequences of Sulfhydryl Modification in the Pore-Forming Subunits of Cardiovascular Ca2+ and Na+ Channels
From the Division of Cardiology (N.C., B.O., T.J.K., E.M.), Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Md; the Institut für Pharmakologie und Toxikologie der Technischen Universität München (Germany) (F.H., V.F.); and the Department of Biochemistry and Biophysics and School of Medicine (R.G.K.), University of Pennsylvania, Philadelphia.
Correspondence to Eduardo Marban, MD, PhD, 844 Ross Bldg, The Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Abstract The structure and function of many
cysteine-containing proteins critically depend on the oxidation state
of the sulfhydryl groups. In such proteins, selective modification of
sulfhydryl groups can be used to probe the relation between structure
and function. We examined the effects of sulfhydryl-oxidizing and
-reducing agents on the function of the heterologously expressed
pore-forming subunits of the cloned rabbit smooth muscle L-type
Ca2+ channel and the human cardiac
tetrodotoxin-insensitive Na+ channel. The known sequences
of the channels suggest the presence of three or four cysteine residues
within the putative pores of Ca2+ or Na+
channels, respectively, as well as multiple other cysteines in regions
of unknown function. We determined the effects of sulfhydryl
modification on Ca2+ and Na+ channel
gating and permeation by using the whole-cell and single-channel
variants of the patch-clamp technique. Within 10 minutes of exposure to
2,2'-dithiodipyridine (DTDP, a specific lipophilic oxidizer of
sulfhydryl groups), Ca2+ current was reduced
compared with the control value, with no significant change in the
kinetics and no shift in the current-voltage relations. The effect
could be readily reversed by 1,4-dithiothreitol (an agent that reduces
disulfide bonds). Similar results were obtained by using the
hydrophilic sulfhydryl-oxidizing agent thimerosal. The effects were
Ca2+-channel specific: DTDP induced no changes in
expressed human cardiac Na+ current. Single-channel
Ba2+ current recordings revealed a reduction in open
probability and mean open time by DTDP but no change in single-channel
conductance, implying that the reduction of macroscopic
Ca2+ current reflects changes in gating and not
permeation. In summary, the pore-forming (
1) subunit of
the L-type Ca2+ channel contains functionally
important free sulfhydryl groups that modulate gating. These free
sulfhydryl groups are accessible from the extracellular side by an
aqueous pathway.
Key Words: Ca2+ channels • Na+ channels • cysteine • sulfhydryl oxidation
This article has been cited by other articles:
 |
|
 |
|
  C. Coddou, J. F. Codocedo, S. Li, J. G. Lillo, C. Acuna-Castillo, P. Bull, S. S. Stojilkovic, and J. P. Huidobro-Toro Reactive Oxygen Species Potentiate the P2X2 Receptor Activity through Intracellular Cys430 J. Neurosci., September 30, 2009; 29(39): 12284 - 12291. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. J. S. Griffioen, H. W. Kamendi, C. J. Gorini, E. Bouairi, and D. Mendelowitz Reactive Oxygen Species Mediate Central Cardiorespiratory Network Responses to Acute Intermittent Hypoxia J Neurophysiol, March 1, 2007; 97(3): 2059 - 2066. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Li, X. Li, Y.-L. Li, C.-H. Shao, K. R. Bidasee, and G. J. Rozanski Insulin regulation of glutathione and contractile phenotype in diabetic rat ventricular myocytes Am J Physiol Heart Circ Physiol, March 1, 2007; 292(3): H1619 - H1629. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Zhang, Y. He, D. Tuteja, D. Xu, V. Timofeyev, Q. Zhang, K. A. Glatter, Y. Xu, H.-S. Shin, R. Low, et al. Functional Roles of Cav1.3({alpha}1D) Calcium Channels in Atria: Insights Gained From Gene-Targeted Null Mutant Mice Circulation, September 27, 2005; 112(13): 1936 - 1944. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. T. Mallet Hypoxic modulation of cardiac L-type Ca2+ current: Interaction of reactive oxygen species and {beta}-adrenergic signaling Cardiovasc Res, September 1, 2005; 67(4): 578 - 580. [Full Text] [PDF]
|
|
|
|
 |
|
 F. Simon, D. Varela, A. L. Eguiguren, L. F. Diaz, F. Sala, and A. Stutzin Hydroxyl radical activation of a Ca2+-sensitive nonselective cation channel involved in epithelial cell necrosis Am J Physiol Cell Physiol, October 1, 2004; 287(4): C963 - C970. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L Alvarez, E. Salinas-Stefanon, G. Orta, T. Ferrer, K. Talavera, L. Galan, and G. Vassort Occurrence of a tetrodotoxin-sensitive calcium current in rat ventricular myocytes after long-term myocardial infarction Cardiovasc Res, September 1, 2004; 63(4): 653 - 661. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. S. Bandali, M. P. Belanger, and C. Wittnich Hyperoxia causes oxygen free radical-mediated membrane injury and alters myocardial function and hemodynamics in the newborn Am J Physiol Heart Circ Physiol, August 1, 2004; 287(2): H553 - H559. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Wang, J. Anrather, J. Huang, R. C. Speth, V. M. Pickel, and C. Iadecola NADPH Oxidase Contributes to Angiotensin II Signaling in the Nucleus Tractus Solitarius J. Neurosci., June 16, 2004; 24(24): 5516 - 5524. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. E. Berry and J. M. Hare Xanthine oxidoreductase and cardiovascular disease: molecular mechanisms and pathophysiological implications J. Physiol., March 15, 2004; 555(3): 589 - 606. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. C. Hool Differential regulation of the slow and rapid components of guinea-pig cardiac delayed rectifier K+ channels by hypoxia J. Physiol., February 1, 2004; 554(3): 743 - 754. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. Gupte, M. Arshad, S. Viola, P. M. Kaminski, Z. Ungvari, G. Rabbani, A. Koller, and M. S. Wolin Pentose phosphate pathway coordinates multiple redox-controlled relaxing mechanisms in bovine coronary arteries Am J Physiol Heart Circ Physiol, December 1, 2003; 285(6): H2316 - H2326. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Zhang, Y. Xu, P. H. Dong, D. Sharma, and N. Chiamvimonvat A negatively charged residue in the outer mouth of rat sodium channel determines the gating kinetics of the channel Am J Physiol Cell Physiol, May 1, 2003; 284(5): C1247 - C1254. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Zhang, Y. Xu, H. Song, J. Rodriguez, D. Tuteja, Y. Namkung, H.-S. Shin, and N. Chiamvimonvat Functional Roles of Cav1.3 ({alpha}1D) Calcium Channel in Sinoatrial Nodes: Insight Gained Using Gene-Targeted Null Mutant Mice Circ. Res., May 17, 2002; 90(9): 981 - 987. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. Gupte, K.-X. Li, T. Okada, K. Sato, and M. Oka Inhibitors of Pentose Phosphate Pathway Cause Vasodilation: Involvement of Voltage-Gated Potassium Channels J. Pharmacol. Exp. Ther., April 1, 2002; 301(1): 299 - 305. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. F. Saavedra, N. Paolocci, M. E. St. John, M. W. Skaf, G. C. Stewart, J.-S. Xie, R. W. Harrison, J. Zeichner, D. Mudrick, E. Marban, et al. Imbalance Between Xanthine Oxidase and Nitric Oxide Synthase Signaling Pathways Underlies Mechanoenergetic Uncoupling in the Failing Heart Circ. Res., February 22, 2002; 90(3): 297 - 304. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T Smani, A Hernandez, J Urena, A.G Castellano, A Franco-Obregon, A Ordonez, and J Lopez-Barneo Reduction of Ca2+ channel activity by hypoxia in human and porcine coronary myocytes Cardiovasc Res, January 1, 2002; 53(1): 97 - 104. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. C. Hool Hypoxia Increases the Sensitivity of the L-Type Ca2+ Current to {beta}-Adrenergic Receptor Stimulation via a C2 Region-Containing Protein Kinase C Isoform Circ. Res., December 8, 2000; 87(12): 1164 - 1171. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Iesaki and M. S. Wolin Thiol Oxidation Activates a Novel Redox-Regulated Coronary Vasodilator Mechanism Involving Inhibition of Ca2+ Influx Arterioscler Thromb Vasc Biol, November 1, 2000; 20(11): 2359 - 2365. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Yatani and T. J. Kamp Tails of the L-Type Ca2+ Channel : To Sense Oxygen or Not Circ. Res., September 29, 2000; 87(7): 535 - 536. [Full Text] [PDF]
|
|
|
|
|
|
O. M. Hernandez, D. J. Discher, N. H. Bishopric, and K. A. Webster Rapid Activation of Neutral Sphingomyelinase by Hypoxia-Reoxygenation of Cardiac Myocytes Circ. Res., February 4, 2000; 86(2): 198 - 204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. E. G. Ekelund, R. W. Harrison, O. Shokek, R. N. Thakkar, R. S. Tunin, H. Senzaki, D. A. Kass, E. Marban, and J. M. Hare Intravenous Allopurinol Decreases Myocardial Oxygen Consumption and Increases Mechanical Efficiency in Dogs With Pacing-Induced Heart Failure Circ. Res., September 3, 1999; 85(5): 437 - 445. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Carmeliet Cardiac Ionic Currents and Acute Ischemia: From Channels to Arrhythmias Physiol Rev, July 1, 1999; 79(3): 917 - 1017. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. G. Tsushima, A. D. Wickenden, R. A. Bouchard, G. Y. Oudit, P. P. Liu, and P. H. Backx Modulation of Iron Uptake in Heart by L-Type Ca2+ Channel Modifiers : Possible Implications in Iron Overload Circ. Res., June 11, 1999; 84(11): 1302 - 1309. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Macháty, W.-H. Wang, B. N. Day, and R. S. Prather Calcium Release and Subsequent Development Induced by Modification of Sulfhydryl Groups in Porcine Oocytes Biol Reprod, June 1, 1999; 60(6): 1384 - 1391. [Abstract] [Full Text]
|
|
|
|
 |
|
 J. Xia, J. D. Browning, and B. L. O'Dell Decreased Plasma Membrane Thiol Concentration Is Associated with Increased Osmotic Fragility of Erythrocytes in Zinc-Deficient Rats J. Nutr., April 1, 1999; 129(4): 814 - 819. [Abstract] [Full Text]
|
|
|
|
|
|
I M Fearon, A C V Palmer, A J Balmforth, S G Ball, G Varadi, and C Peers Modulation of recombinant human cardiac L-type Ca2+ channel {alpha}1C subunits by redox agents and hypoxia J. Physiol., February 1, 1999; 514(3): 629 - 637. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Zucchi, G. Yu, P. Galbani, M. Mariani, G. Ronca, and S. Ronca-Testoni Sulfhydryl Redox State Affects Susceptibility to Ischemia and Sarcoplasmic Reticulum Ca2+ Release in Rat Heart : Implications for Ischemic Preconditioning Circ. Res., November 2, 1998; 83(9): 908 - 915. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Li, J. Segui, S. H. Heinemann, and T. Hoshi Oxidation Regulates Cloned Neuronal Voltage-Dependent Ca2+ Channels Expressed in Xenopus Oocytes J. Neurosci., September 1, 1998; 18(17): 6740 - 6747. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. I. Kourie Interaction of reactive oxygen species with ion transport mechanisms Am J Physiol Cell Physiol, July 1, 1998; 275(1): C1 - C24. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J Taggart and S. Wray Hypoxia and smooth muscle function: key regulatory events during metabolic stress J. Physiol., June 1, 1998; 509(2): 315 - 325. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Xu, J. P. Eu, G. Meissner, and J. S. Stamler Activation of the Cardiac Calcium Release Channel (Ryanodine Receptor) by Poly-S-Nitrosylation Science, January 9, 1998; 279(5348): 234 - 237. [Abstract] [Full Text]
|
|
|
|
|
|
H. Hu Nipavan Chiamvimonvat Toshio Yamagishi Eduardo Direct Inhibition of Expressed Cardiac L-Type Ca2+ Channels by S-Nitrosothiol Nitric Oxide Donors Circ. Res., November 19, 1997; 81(5): 742 - 752. [Abstract] [Full Text]
|
|
|
|
|
|
T. J. DiChiara and P. H. Reinhart Redox Modulation of hslo Ca2+-Activated K+ Channels J. Neurosci., July 1, 1997; 17(13): 4942 - 4955. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Tritto, D. D'Andrea, N. Eramo, A. Scognamiglio, C. De Simone, A. Violante, A. Esposito, M. Chiariello, and G. Ambrosio Oxygen Radicals Can Induce Preconditioning in Rabbit Hearts Circ. Res., May 19, 1997; 80(5): 743 - 748. [Abstract] [Full Text]
|
|
|
|
|
|
R. A. Kelly, J.-L. Balligand, and T. W. Smith Nitric Oxide and Cardiac Function Circ. Res., September 1, 1996; 79(3): 363 - 380. [Full Text]
|
|
|
|
 |
|
 D. R. Meldrum, J. C. Cleveland Jr, B. C. Sheridan, R. T. Rowland, A. Banerjee, and A. H. Harken Cardiac Surgical Implications of Calcium Dyshomeostasis in the Heart Ann. Thorac. Surg., April 1, 1996; 61(4): 1273 - 1280. [Abstract] [Full Text]
|
|
|
|
|
|
W. Chen, S. Gabel, C. Steenbergen, and E. Murphy A Redox-Based Mechanism for Cardioprotection Induced by Ischemic Preconditioning in Perfused Rat Heart Circ. Res., August 1, 1995; 77(2): 424 - 429. [Abstract] [Full Text]
|
|
|
|
 |
|
 X.-S. Wu, H. D. Edwards, and W. A. Sather Side Chain Orientation in the Selectivity Filter of a Voltage-gated Ca2+ Channel J. Biol. Chem., October 6, 2000; 275(41): 31778 - 31785. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. F. Saavedra, N. Paolocci, M. E. St. John, M. W. Skaf, G. C. Stewart, J.-S. Xie, R. W. Harrison, J. Zeichner, D. Mudrick, E. Marban, et al. Imbalance Between Xanthine Oxidase and Nitric Oxide Synthase Signaling Pathways Underlies Mechanoenergetic Uncoupling in the Failing Heart Circ. Res., February 22, 2002; 90(3): 297 - 304. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Forbes, C. Steenbergen, and E. Murphy Diazoxide-Induced Cardioprotection Requires Signaling Through a Redox-Sensitive Mechanism Circ. Res., April 27, 2001; 88(8): 802 - 809. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. U. Ahmmed, Y. Xu, P. Hong Dong, Z. Zhang, J. Eiserich, and N. Chiamvimonvat Nitric Oxide Modulates Cardiac Na+ Channel via Protein Kinase A and Protein Kinase G Circ. Res., November 23, 2001; 89(11): 1005 - 1013. [Abstract] [Full Text] [PDF]
|
|