scholarly journals Antibody probe study of Ca2+ channel regulation by interdomain interaction within the ryanodine receptor

2004 ◽  
Vol 380 (2) ◽  
pp. 561-569 ◽  
Author(s):  
Shigeki KOBAYASHI ◽  
Takeshi YAMAMOTO ◽  
Jerome PARNESS ◽  
Noriaki IKEMOTO
Keyword(s):  
Ryanodine Receptor ◽  
Fluorescence Probe ◽  
Aqueous Environment ◽  
Channel Activation ◽  
Regulatory Domains ◽  
Domain Specific ◽  
Terminal Domain ◽  
Domain Interactions ◽  
Interdomain Interaction ◽  
Domain Peptide

N-terminal and central domains of ryanodine receptor 1 (RyR1), where many reported malignant hyperthermia (MH) mutations are localized, represent putative channel regulatory domains. Recent domain peptide (DP) probe studies led us to the hypothesis that these domains interact to stabilize the closed state of channel (zipping), while weakening of domain–domain interactions (unzipping) by mutation de-stabilizes the channel, making it leaky to Ca2+ or sensitive to the agonists of RyR1. As shown previously, DP1 (N-terminal domain peptide) and DP4 (central domain peptide) produced MH-like channel activation/sensitization effects, presumably by peptide binding to sites critical to stabilizing domain–domain interactions and resultant loss of conformational constraints. Here we report that polyclonal anti-DP1 and anti-DP4 antibodies also produce MH-like channel activation and sensitization effects as evidenced by about 4-fold enhancement of high affinity [3H]ryanodine binding to RyR1 and by a significant left-shift of the concentration-dependence of activation of sarcoplasmic reticulum Ca2+ release by polylysine. Fluorescence quenching experiments demonstrate that the accessibility of a DP4-directed, conformationally sensitive fluorescence probe linked to the RyR1 N-terminal domain is increased in the presence of domain-specific antibodies, consistent with the view that these antibodies produce unzipping of interacting domains that are of hindered accessibility to the surrounding aqueous environment. Our results suggest that domain-specific antibody binding induces a conformational change resulting in channel activation, and are consistent with the hypothesis that interacting N-terminal and central domains are intimately involved in the regulation of RyR1 channel function.

scholarly journals Malignant hyperthermia mutation sites in the Leu2442–Pro2477 (DP4) region of RyR1 (ryanodine receptor 1) are clustered in a structurally and functionally definable area

2006 ◽  
Vol 401 (1) ◽  
pp. 333-339 ◽  
Author(s):  
Mark L. Bannister ◽  
Tomoyo Hamada ◽  
Takashi Murayama ◽  
Peta J. Harvey ◽  
Marco G. Casarotto ◽  
...  
Keyword(s):  
Malignant Hyperthermia ◽  
Ryanodine Receptor ◽  
Structural Studies ◽  
Amino Acid Residues ◽  
Domain Interaction ◽  
Closed State ◽  
Interdomain Interaction ◽  
Ryanodine Receptor 1 ◽  
Domain Peptide ◽  
Interdomain Interactions

To explain the mechanism of pathogenesis of channel disorder in MH (malignant hyperthermia), we have proposed a model in which tight interactions between the N-terminal and central domains of RyR1 (ryanodine receptor 1) stabilize the closed state of the channel, but mutation in these domains weakens the interdomain interaction and destabilizes the channel. DP4 (domain peptide 4), a peptide corresponding to residues Leu2442–Pro2477 of the central domain, also weakens the domain interaction and produces MH-like channel destabilization, whereas an MH mutation (R2458C) in DP4 abolishes these effects. Thus DP4 and its mutants serve as excellent tools for structure–function studies. Other MH mutations have been reported in the literature involving three other amino acid residues in the DP4 region (Arg2452, Ile2453 and Arg2454). In the present paper we investigated the activity of several mutants of DP4 at these three residues. The ability to activate ryanodine binding or to effect Ca2+ release was severely diminished for each of the MH mutants. Other substitutions were less effective. Structural studies, using NMR analysis, revealed that the peptide has two α-helical regions. It is apparent that the MH mutations are clustered at the C-terminal end of the first helix. The data in the present paper indicates that mutation of residues in this region disrupts the interdomain interactions that stabilize the closed state of the channel.

Site-specific modification of calmodulin Ca2+ affinity tunes the skeletal muscle ryanodine receptor activation profile

2010 ◽  
Vol 432 (1) ◽  
pp. 89-99 ◽  
Author(s):  
Jie Jiang ◽  
Yubin Zhou ◽  
Jin Zou ◽  
Yanyi Chen ◽  
Priya Patel ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Binding Affinity ◽  
Ryanodine Receptor ◽  
Receptor Activation ◽  
Release Channel ◽  
Domain Specific ◽  
Terminal Domain ◽  
Channel Opening ◽  
Activation Profile ◽  
Ef Hands

The skeletal muscle isoform of the ryanodine receptor Ca2+-release channel (RyR1) is regulated by Ca2+ and CaM (calmodulin). CaM shifts the biphasic Ca2+-dependence of RyR1 activation leftward, effectively increasing channel opening at low Ca2+ and decreasing channel opening at high Ca2+. The conversion of CaM from a RyR1 activator into an inhibitor is due to the binding of Ca2+ to CaM; however, which of CaM's four Ca2+-binding sites serves as the switch for this conversion is unclear. We engineered a series of mutant CaMs designed to individually increase the Ca2+ affinity of each of CaM's EF-hands by increasing the number of acidic residues in Ca2+-chelating positions. Domain-specific Ca2+ affinities of each CaM variant were determined by equilibrium fluorescence titration. Mutations in sites I (T26D) or II (N60D) in CaM's N-terminal domain had little effect on CaM Ca2+ affinity and regulation of RyR1. However, the site III mutation N97D increased the Ca2+-binding affinity of CaM's C-terminal domain and caused CaM to inhibit RyR1 at a lower Ca2+ concentration than wild-type CaM. Conversely, the site IV mutation Q135D decreased the Ca2+-binding affinity of CaM's C-terminal domain and caused CaM to inhibit RyR1 at higher Ca2+ concentrations. These results support the hypothesis that Ca2+ binding to CaM's C-terminal acts as the switch converting CaM from a RyR1 activator into a channel inhibitor. These results indicate further that targeting CaM's Ca2+ affinity may be a valid strategy to tune the activation profile of CaM-regulated ion channels.

Crystal Structure of the N-terminal Domain of Ryanodine Receptor from Plutella xylostella

10.3791/58568 ◽  
2018 ◽  
Author(s):  
Bidhan Chandra Nayak ◽  
Jie Wang ◽  
Lianyun Lin ◽  
Weiyi He ◽  
Minsheng You ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Ryanodine Receptor ◽  
Plutella Xylostella ◽  
Terminal Domain

scholarly journals The N-Terminal Region of the Ryanodine Receptor Affects Channel Activation

2017 ◽  
Vol 8 ◽  
Author(s):  
Andrea Faltinova ◽  
Nataša Tomaskova ◽  
Marián Antalik ◽  
Jozef Sevcik ◽  
Alexandra Zahradnikova
Keyword(s):  
Ryanodine Receptor ◽  
Terminal Region ◽  
Channel Activation

scholarly journals Substrate Specificity of the FurE Transporter Is Determined by Cytoplasmic Terminal Domain Interactions

Genetics ◽  
2017 ◽  
pp. genetics.300327.2017 ◽  
Author(s):  
Georgia F. Papadaki ◽  
Sotiris Amillis ◽  
George Diallinas
Keyword(s):  
Substrate Specificity ◽  
Terminal Domain ◽  
Domain Interactions

scholarly journals The arrhythmogenic N53I variant subtly changes the structure and dynamics in the calmodulin N-terminal domain, altering its interaction with the cardiac ryanodine receptor

2020 ◽  
Vol 295 (22) ◽  
pp. 7620-7634
Author(s):  
Christian Holt ◽  
Louise Hamborg ◽  
Kelvin Lau ◽  
Malene Brohus ◽  
Anders Bundgaard Sørensen ◽  
...  
Keyword(s):  
Ventricular Tachycardia ◽  
Ryanodine Receptor ◽  
Hydrophobic Surface ◽  
Catecholaminergic Polymorphic Ventricular Tachycardia ◽  
Polymorphic Ventricular Tachycardia ◽  
Open Probability ◽  
Calmodulin Binding ◽  
Terminal Domain ◽  
Genes Encoding ◽  
Cardiac Ryanodine Receptor

Mutations in the genes encoding the highly conserved Ca2+-sensing protein calmodulin (CaM) cause severe cardiac arrhythmias, including catecholaminergic polymorphic ventricular tachycardia or long QT syndrome and sudden cardiac death. Most of the identified arrhythmogenic mutations reside in the C-terminal domain of CaM and mostly affect Ca2+-coordinating residues. One exception is the catecholaminergic polymorphic ventricular tachycardia–causing N53I substitution, which resides in the N-terminal domain (N-domain). It does not affect Ca2+ coordination and has only a minor impact on binding affinity toward Ca2+ and on other biophysical properties. Nevertheless, the N53I substitution dramatically affects CaM's ability to reduce the open probability of the cardiac ryanodine receptor (RyR2) while having no effect on the regulation of the plasmalemmal voltage-gated Ca2+ channel, Cav1.2. To gain more insight into the molecular disease mechanism of this mutant, we used NMR to investigate the structures and dynamics of both apo- and Ca2+-bound CaM-N53I in solution. We also solved the crystal structures of WT and N53I CaM in complex with the primary calmodulin-binding domain (CaMBD2) from RyR2 at 1.84–2.13 Å resolutions. We found that all structures of the arrhythmogenic CaM-N53I variant are highly similar to those of WT CaM. However, we noted that the N53I substitution exposes an additional hydrophobic surface and that the intramolecular dynamics of the protein are significantly altered such that they destabilize the CaM N-domain. We conclude that the N53I-induced changes alter the interaction of the CaM N-domain with RyR2 and thereby likely cause the arrhythmogenic phenotype of this mutation.

Incorporation of an Unnatural Amino Acid as a Domain-Specific Fluorescence Probe in a Two-Domain Protein

Biochemistry ◽  
2016 ◽  
Vol 55 (49) ◽  
pp. 6739-6742 ◽  
Author(s):  
Lena K. Ries ◽  
Franz X. Schmid ◽  
Philipp A. M. Schmidpeter
Keyword(s):  
Amino Acid ◽  
Fluorescence Probe ◽  
Unnatural Amino Acid ◽  
Specific Fluorescence ◽  
Domain Specific ◽  
Domain Protein

scholarly journals Yeast PIAS-type Ull1/Siz1 Is Composed of SUMO Ligase and Regulatory Domains

2005 ◽  
Vol 280 (43) ◽  
pp. 35822-35828 ◽  
Author(s):  
Yoshimitsu Takahashi ◽  
Yoshiko Kikuchi
Keyword(s):  
Neck Region ◽  
Binding Motif ◽  
Conjugation System ◽  
Regulatory Domains ◽  
Terminal Domain ◽  
Sumo Ligase ◽  
Lysine Residues ◽  
Protein Instability ◽  
Two Hybrid

SUMO (small ubiquitin-like modifier)/Smt3 (suppressor of mif two) is a member of the ubiquitin-related protein family and is known to conjugate with many proteins. In the sumoylation pathway, SUMO/Smt3 is transferred to substrate lysine residues through the thioester cascade of E1 (activating enzyme) and E2 (conjugating enzyme), and E3 (SUMO ligase) functions as an adaptor between E2 and each substrate. Yeast Ull1 (ubiquitin-like protein ligase 1)/Siz1, a PIAS (protein inhibitor of activated STAT)-type SUMO ligase, modifies both cytoplasmic and nuclear proteins. In this paper, we performed a domain analysis of Ull1/Siz1 by constructing various deletion mutants. A novel conserved N-terminal domain, called PINIT, as well as the RING-like domain (SP-RING) were required for the SUMO ligase activity in the in vitro conjugation system and for interaction with Smt3 in an in vitro binding assay. The most distal N-terminal region, which contains a putative DNA-binding SAF-A/B, Acinus, and PIAS (SAP) motif, was not required for the ligase activity but was involved in nuclear localization. A strong SUMO-binding motif was identified, which interacted with Smt3 in the two-hybrid system but was not necessary for the ligase activity. The most distal C-terminal domain was important for stable localization at the bud neck region and thereby for the substrate recognition of septins. Furthermore, the C-terminal half conferred protein instability on Ull1/Siz1. Taken together, we conclude that the SP-RING and PINIT of Ull1/Siz1 are core domains of the SUMO ligase, and the other domains are regulatory for protein stability and subcellular localization.

Bi-directional N-terminal and Central Domain Interactions as a Critical Mechanism for Channel Gating of Ryanodine Receptor

2006 ◽  
Vol 12 (8) ◽  
pp. S164
Author(s):  
Hiroki Tateishi ◽  
Masafumi Yano ◽  
Takeshi Yamamoto ◽  
Xiaojuan Xu ◽  
Hitoshi Uchinoumi ◽  
...  
Keyword(s):  
Ryanodine Receptor ◽  
Channel Gating ◽  
Central Domain ◽  
Domain Interactions

scholarly journals Calcium-dependent energetics of calmodulin domain interactions with regulatory regions of the Ryanodine Receptor Type 1 (RyR1)

2014 ◽  
Vol 193-194 ◽  
pp. 35-49 ◽  
Author(s):  
Rhonda A. Newman ◽  
Brenda R. Sorensen ◽  
Adina M. Kilpatrick ◽  
Madeline A. Shea
Keyword(s):  
Ryanodine Receptor ◽  
Receptor Type ◽  
Regulatory Regions ◽  
Calcium Dependent ◽  
Domain Interactions
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