Bastadins relate ryanodine-sensitive and -insensitive Ca2+ efflux pathways in skeletal SR and BC3H1 cells

Isaac N Pessah, Tadeusz F. Molinski, Trena D. Meloy, Patty Wong, Edmond D. Buck, Paul D. Allen, Frederick C Mohr, Matthew M. Mack

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Bastadins potently interact with the FK-506-binding protein of 12 kDa (FKBP12)-ryanodine receptor (Ry1R) complex in skeletal muscle to enhance a high-affinity ryanodine binding conformation (M. M. Mack, T. F. Molinski, E. D. Buck, and I. N. Pessah. J. Biol. Chem. 269: 23236-23249, 1994). Bastadins are used to examine the relationship between ryanodine-sensitive and ryanodine-insensitive Ca2+ efflux pathways that coexist in junctional sarcoplasmic reticulum (SR) vesicles from rabbit skeletal muscle and differentiated BC3H1 cells. Complete block of caffeine-sensitive Ca2+ channels with micromolar ryanodine or ruthenium red does not alter the steady-state loading capacity of SR. Inhibition of sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) pumps with thapsigargin unmasks a ryanodine- and ruthenium red-insensitive Ca2+ efflux pathway. Bastadin 5 alone does not inhibit Ca2+ efflux unmasked by inhibition of SERCA pumps, but, in combination with blocking concentrations of ryanodine or ruthenium red, it eliminates the ryanodine-insensitive Ca2+ 'leak' and enhances steady-state loading capacity of SR vesicles ~2.5-fold. These actions of bastadins occur in the same concentration range that enhances the number of high-affinity binding sites for [3H]ryanodine (50% effective concentration of ~2 μM). Similar effects on SR Ca2+ transport are found with FK-506 and ryanodine in combination. Block of Ry1R in intact BC3H1 cells with ryanodine does not eliminate the prominent Ca2+ leak unmasked by thapsigargin. A membrane- permeant mixture of bastadins in combination with ryanodine nearly eliminates the Ca2+ leak unmasked by thapsigargin, even though the Ca2+ stores are replete. The requirement of both a known Ry1R blocker and bastadins in combination provides a pharmacological link between ryanodine-sensitive Ca2+ channels and ryanodine-insensitive leak pathways in isolated junctional SR and BC3H1 cells. Together, these results strongly suggest that bastadins, through their modulatory actions on the FKBP12-Ry1R complex, convert ryanodine-insensitive leak states into ryanodine-sensitive channels that recognize [3H]ryanodine with high affinity.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume272
Issue number2 41-2
StatePublished - Feb 1997

Fingerprint

Ryanodine
Sarcoplasmic Reticulum
Ruthenium Red
Thapsigargin
Tacrolimus Binding Proteins
Reticulum
Calcium-Transporting ATPases
Muscle
Skeletal Muscle
Pumps
Ryanodine Receptor Calcium Release Channel
Tacrolimus
Caffeine

Keywords

  • FKBP12
  • ryanodine receptor
  • sarcoplasmic reticulum
  • sarcoplasmic reticulum calcium channels
  • sarcoplasmic reticulum calcium leak

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

Pessah, I. N., Molinski, T. F., Meloy, T. D., Wong, P., Buck, E. D., Allen, P. D., ... Mack, M. M. (1997). Bastadins relate ryanodine-sensitive and -insensitive Ca2+ efflux pathways in skeletal SR and BC3H1 cells. American Journal of Physiology - Cell Physiology, 272(2 41-2).

Bastadins relate ryanodine-sensitive and -insensitive Ca2+ efflux pathways in skeletal SR and BC3H1 cells. / Pessah, Isaac N; Molinski, Tadeusz F.; Meloy, Trena D.; Wong, Patty; Buck, Edmond D.; Allen, Paul D.; Mohr, Frederick C; Mack, Matthew M.

In: American Journal of Physiology - Cell Physiology, Vol. 272, No. 2 41-2, 02.1997.

Research output: Contribution to journalArticle

Pessah, Isaac N ; Molinski, Tadeusz F. ; Meloy, Trena D. ; Wong, Patty ; Buck, Edmond D. ; Allen, Paul D. ; Mohr, Frederick C ; Mack, Matthew M. / Bastadins relate ryanodine-sensitive and -insensitive Ca2+ efflux pathways in skeletal SR and BC3H1 cells. In: American Journal of Physiology - Cell Physiology. 1997 ; Vol. 272, No. 2 41-2.
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