A Procedure for Purification of the Ryanodine Receptor from Skeletal Muscle

Hawkes, Mary Jane; Dı́az-Muñoz, Mauricio; Hamilton, Susan L.

doi:10.3109/09687688909025827

Cited by 39 publications

(21 citation statements)

References 18 publications

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“…2A). Similar behavior of the solubilized RyR has been previously described (14,33). In contrast, analysis of the sedimentation profile of the DHP-R (Fig.…”

Section: Resultssupporting

confidence: 85%

“…In addition to the major polypeptide, a minor band of 400 kDa was recognized by the anti-RyR antibodies. This peptide corresponds to the degradation product of the RyR already described by other groups (15,32,33), as confirmed by the increase in the intensity of the corresponding band in absence of protease inhibitors and by the presence of this peptide in purified RyR Proc. Natl.…”

Section: Resultssupporting

confidence: 78%

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Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle.

Marty

Robert

Villaz

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

145

123

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Membrane vesicles enriched in both ryanodine receptor and dihydropyridine receptor were obtained from rabbit skeletal muscle and solubilized with 3- [(3-cholamidopropyl) (14,17,18).Three different mechanisms have been proposed for excitation-contraction coupling in skeletal muscle: Ca2 -induced Ca2+ release, inositol 1,4,5-trisphosphate-induced Ca2W release, and direct physical coupling (for review see refs. 19 and 20). Current evidence favors a model in which a voltagedriven conformational change ofthe al subunit ofthe DHP-R activates Ca2+ release by the RyR through direct physical interaction (11,21,22

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“…2A). Similar behavior of the solubilized RyR has been previously described (14,33). In contrast, analysis of the sedimentation profile of the DHP-R (Fig.…”

Section: Resultssupporting

confidence: 85%

Section: Resultssupporting

confidence: 78%

Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle.

Marty

Robert

Villaz

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

145

123

View full text Add to dashboard Cite

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“…Skeletal muscle membranes were prepared from rabbit fast-twitch skeletal muscle, as described in Hawkes et al (17). Homogeneous preparations of the skeletal muscle Ca 2ϩ channel suitable for studies by electron microscopy were purified by using WGA-Affi-Gel column and DEAE-Trisacryl M ion exchange as described (18) with some modifications to adapt this procedure for structural studies.…”

Section: Methodsmentioning

confidence: 99%

Structure of the voltage-gated L-type Ca ²⁺ channel by electron cryomicroscopy

Serysheva

Ludtke

Baker

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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Voltage-dependent L-type Ca 2؉ channels play important functional roles in many excitable cells. We present a three-dimensional structure of an L-type Ca 2؉ channel. Electron cryomicroscopy in conjunction with single-particle processing was used to determine a 30-Å resolution structure of the channel protein. The asymmetrical channel structure consists of two major regions: a heartshaped region connected at its widest end with a handle-shaped region. A molecular model is proposed for the arrangement of this skeletal muscle L-type Ca 2؉ channel structure with respect to the sarcoplasmic reticulum Ca 2؉ -release channel, the physical partner of the L-type channel for signal transduction during the excitationcontraction coupling in muscle.

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“…SR Membrane Preparation-SR and transverse tubule-enriched membranes were prepared from rabbit hind leg and back-strap skeletal muscle and purified using sucrose gradient centrifugation (19). Protein concentrations were estimated by the method of Lowry using bovine serum albumin as the standard (20).…”

Section: Methodsmentioning

confidence: 99%

Suramin Interacts with the Calmodulin Binding Site on the Ryanodine Receptor, RYR1

Papineni¹,

O’Connell²,

Zhang³

et al. 2002

Journal of Biological Chemistry

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Apocalmodulin and Ca2؉ calmodulin bind to overlapping sites on the ryanodine receptor skeletal form, RYR1, but have opposite functional effects on channel activity. Suramin, a polysulfonated napthylurea, displaces both forms of calmodulin, leading to an inhibition of activity at low Ca 2؉ The skeletal muscle ryanodine receptor (RYR1) functions as a sarcoplasmic reticulum (SR) 1 Ca 2ϩ release channel that plays a central role in excitation-contraction coupling. Two distinct mechanisms are postulated to contribute to the release of Ca 2ϩ from the SR in skeletal muscle. After sarcolemmal depolarization, SR Ca 2ϩ release channels are initially activated via mechanical coupling with DHPRs (or L-type Ca 2ϩ channels) located in the surface membrane (2). However, morphological data indicate that only every other SR Ca 2ϩ release channel is directly coupled with sarcolemmal DHPRs (3). Adjacent, non-DHPR-coupled release channels (4) are thought to be activated either by Ca 2ϩ released via the mechanically coupled channels (3) or by a coordinated or "coupled-gating" mechanism of activation (5).RYR1 and DHPR proteins bind CaM in both its Ca 2ϩ -bound and Ca 2ϩ -free forms (6 -9). Overlapping binding sites for apoCaM and Ca 2ϩ -CaM are located between amino acids 3614 and 3643 of RYR1 (10, 11). The carboxyl-terminal tail of the DHPR ␣ 1 -subunit (12, 13) appears to have binding sites for both forms of CaM. C3635, located within the putative CaM binding region of RYR1, has been postulated to contribute to oxidation-induced intersubunit cross-linking (14) and was recently demonstrated to be the site of CaM-dependent NO modulation of RYR1 (15).Studies of the interaction of both the DHPR and RYR1 with apoCaM and Ca 2ϩ CaM have been primarily carried out with uncoupled channels, raising the question of whether CaM interacts with either channel when the two proteins are mechanically coupled in intact skeletal muscle. Slavik et al. (12) provided evidence that a sequence within the carboxyl terminus of the DHPR ␣ 1 -subunit interacts strongly with RYR1. This sequence was later shown to also be a CaM binding motif (13). More recently, Sencer et al. (1) demonstrated that the CaM binding site on RYR1 binds directly to the carboxyl-terminal tail of the DHPR ␣ 1 -subunit. These findings suggest that the CaM binding motifs on the DHPR and RYR1 proteins may actually function as protein-protein interaction motifs rather than strictly as CaM binding domains. However, the functionally relevant binding partners for these motifs have not yet been identified. If the DHPR and RYR1 proteins utilize CaM binding motifs for binding to one another, then CaM could potentially uncouple the mechanical link formed between the CaM binding site of RYR1 and the carboxyl-terminal tail of the DHPR ␣ 1 -subunit. However, the functional consequences of such CaM-mediated uncoupling have yet to be evaluated.Suramin, a polysulfonated napthylurea is a potent, reversible activator of the RYR, increasing both conductance and P 0 of the channel (16). Klinger et a...

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A Procedure for Purification of the Ryanodine Receptor from Skeletal Muscle

Cited by 39 publications

References 18 publications

Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle.

Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle.

Structure of the voltage-gated L-type Ca ²⁺ channel by electron cryomicroscopy

Suramin Interacts with the Calmodulin Binding Site on the Ryanodine Receptor, RYR1

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A Procedure for Purification of the Ryanodine Receptor from Skeletal Muscle

Cited by 39 publications

References 18 publications

Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle.

Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle.

Structure of the voltage-gated L-type Ca 2+ channel by electron cryomicroscopy

Suramin Interacts with the Calmodulin Binding Site on the Ryanodine Receptor, RYR1

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Structure of the voltage-gated L-type Ca ²⁺ channel by electron cryomicroscopy