Structural and functional characterization of the purified cardiac ryanodine receptor-Ca2+ release channel complex - PubMed (original) (raw)
. 1989 Jan 15;264(2):1329-35.
Affiliations
- PMID: 2463249
Free article
Structural and functional characterization of the purified cardiac ryanodine receptor-Ca2+ release channel complex
K Anderson et al. J Biol Chem. 1989.
Free article
Abstract
Using density gradient centrifugation and [3H]ryanodine as a specific marker, the ryanodine receptor-Ca2+ release channel complex from Chaps-solubilized canine cardiac sarcoplasmic reticulum (SR) has been purified in the form of an approximately 30 S complex, comprised of Mr approximately 400,000 polypeptides. Purification resulted in a specific activity of approximately 450 pmol bound ryanodine/mg of protein, a 60-70% recovery of ryanodine binding activity, and retention of the high affinity ryanodine binding site (KD = 3 nM). Negative stain electron microscopy revealed a 4-fold symmetric, four-leaf clover structure, which could fill a box approximately 30 x 30 nm and was thus morphologically similar to the SR-transverse-tubule, junctionally associated foot structure. The structural, sedimentation, and ryanodine binding data strongly suggest there is one high affinity ryanodine binding site/30 S complex, comprised of four Mr approximately 400,000 subunits. Upon reconstitution into planar lipid bilayers, the purified complex exhibited a Ca2+ conductance (70 pS in 50 mM Ca2+) similar to that of the native cardiac Ca2+ release channel (75 pS). The reconstituted complex was also found to conduct Na+ (550 pS in 500 mM Na+) and often to display complex Na+ subconducting states. The purified channel could be activated by micromolar Ca2+ or millimolar ATP, inhibited by millimolar Mg2+ or micromolar ruthenium red, and modified to a long-lived open subconducting state by ryanodine. The sedimentation, subunit composition, morphological, and ryanodine binding characteristics of the purified cardiac ryanodine receptor-Ca2+ release channel complex were similar to those previously described for the purified ryanodine receptor-Ca2+ release channel complex from fast-twitch skeletal muscle.
Similar articles
- Purified ryanodine receptor from skeletal muscle sarcoplasmic reticulum is the Ca2+-permeable pore of the calcium release channel.
Imagawa T, Smith JS, Coronado R, Campbell KP. Imagawa T, et al. J Biol Chem. 1987 Dec 5;262(34):16636-43. J Biol Chem. 1987. PMID: 2445748 - Characterization of junctional and longitudinal sarcoplasmic reticulum from heart muscle.
Inui M, Wang S, Saito A, Fleischer S. Inui M, et al. J Biol Chem. 1988 Aug 5;263(22):10843-50. J Biol Chem. 1988. PMID: 2968985 - The muscle ryanodine receptor and its intrinsic Ca2+ channel activity.
Lai FA, Meissner G. Lai FA, et al. J Bioenerg Biomembr. 1989 Apr;21(2):227-46. doi: 10.1007/BF00812070. J Bioenerg Biomembr. 1989. PMID: 2546931 Review. - Ion channels in the sarcoplasmic reticulum of striated muscle.
Dulhunty AF, Junankar PR, Eager KR, Ahern GP, Laver DR. Dulhunty AF, et al. Acta Physiol Scand. 1996 Mar;156(3):375-85. doi: 10.1046/j.1365-201X.1996.193000.x. Acta Physiol Scand. 1996. PMID: 8729698 Review.
Cited by
- Cardiac L-type calcium channel regulation by Leucine-Rich Repeat-Containing Protein 10.
Siri-Angkul N, Kamp TJ. Siri-Angkul N, et al. Channels (Austin). 2024 Dec;18(1):2355121. doi: 10.1080/19336950.2024.2355121. Epub 2024 May 19. Channels (Austin). 2024. PMID: 38762910 Free PMC article. Review. - The structural basis of ryanodine receptor ion channel function.
Meissner G. Meissner G. J Gen Physiol. 2017 Dec 4;149(12):1065-1089. doi: 10.1085/jgp.201711878. Epub 2017 Nov 9. J Gen Physiol. 2017. PMID: 29122978 Free PMC article. Review. - Ultrastructural Analysis of Self-Associated RyR2s.
Cabra V, Murayama T, Samsó M. Cabra V, et al. Biophys J. 2016 Jun 21;110(12):2651-2662. doi: 10.1016/j.bpj.2016.05.013. Biophys J. 2016. PMID: 27332123 Free PMC article. - Type 2 ryanodine receptors are highly sensitive to alcohol.
Ye Y, Jian K, Jaggar JH, Bukiya AN, Dopico AM. Ye Y, et al. FEBS Lett. 2014 May 2;588(9):1659-65. doi: 10.1016/j.febslet.2014.03.005. Epub 2014 Mar 12. FEBS Lett. 2014. PMID: 24631538 Free PMC article. - Mechanisms of altered Ca²⁺ handling in heart failure.
Luo M, Anderson ME. Luo M, et al. Circ Res. 2013 Aug 30;113(6):690-708. doi: 10.1161/CIRCRESAHA.113.301651. Circ Res. 2013. PMID: 23989713 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Research Materials
Miscellaneous