Phosphorylation-dependent Changes in Nucleotide Binding, Conformation, and Dynamics of the First Nucleotide Binding Domain (NBD1) of the Sulfonylurea Receptor 2B (SUR2B)

Araujo, Elvin D. de; Alvarez, Claudia P.; López‐Alonso, Jorge P.; Sooklal, Clarissa R.; Stagljar, Marijana; Kanelis, Voula

doi:10.1074/jbc.m114.636233

Cited by 14 publications

(55 citation statements)

References 97 publications

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“…Thus, it is possible that the C-terminal tail regulates interactions of other regions in gp74 with the terminase enzymes or with DNA. Binding of disordered regions that possess residual secondary structure with folded domains has been observed to affect the activities of other proteins (39,41,82,83). Of course, it is also possible that the C-terminal tail directly binds the terminase enzymes or DNA.…”

Section: Discussionmentioning

confidence: 99%

“…The triple-resonance assignment data were run on a 500-MHz spectrometer equipped with a room-temperature triple-resonance HCN probe with actively shielded z gradients (QANUC NMR Facility, McGill University) at 25°C. The gp74-WT data were supplemented with 15 N-1 H heteronuclear single quantum coherence (HSQC) spectra recorded at 600 MHz on samples that were 15 N labeled only on either Leu or Val (82,83,90). All spectra were referenced with 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS) (91) and processed using NMRPipe/NMRDraw (92).…”

Section: Methodsmentioning

confidence: 99%

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HK97 gp74 Possesses an α-Helical Insertion in the ββα Fold That Affects Its Metal Binding, cos Site Digestion, and In Vivo Activities

et al. 2020

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The last gene in the genome of the bacteriophage HK97 encodes gp74, an HNH endonuclease. HNH motifs contain two conserved His residues and an invariant Asn residue, and they adopt a ββα structure. gp74 is essential for phage head morphogenesis, likely because gp74 enhances the specific endonuclease activity of the HK97 terminase complex. Notably, the ability of gp74 to enhance the terminase-mediated cleavage of the phage cos site requires an intact HNH motif in gp74. Mutation of H82, the conserved metal-binding His residue in the HNH motif, to Ala abrogates gp74-mediated stimulation of terminase activity. Here, we present nuclear magnetic resonance (NMR) studies demonstrating that gp74 contains an α-helical insertion in the Ω-loop, which connects the two β-strands of the ββα fold, and a disordered C-terminal tail. NMR data indicate that the Ω-loop insert makes contacts to the ββα fold and influences the ability of gp74 to bind divalent metal ions. Further, the Ω-loop insert and C-terminal tail contribute to gp74-mediated DNA digestion and to gp74 activity in phage morphogenesis. The data presented here enrich our molecular-level understanding of how HNH endonucleases enhance terminase-mediated digestion of the cos site and contribute to the phage replication cycle. IMPORTANCE This study demonstrates that residues outside the canonical ββα fold, namely, the Ω-loop α-helical insert and a disordered C-terminal tail, regulate the activity of the HNH endonuclease gp74. The increased divalent metal ion binding when the Ω-loop insert is removed compared to reduced cos site digestion and phage formation indicates that the Ω-loop insert plays multiple regulatory roles. The data presented here provide insights into the molecular basis of the involvement of HNH proteins in phage DNA packing.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

HK97 gp74 Possesses an α-Helical Insertion in the ββα Fold That Affects Its Metal Binding, cos Site Digestion, and In Vivo Activities

et al. 2020

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“…SUR2B subunit is mainly responsible for channel affinity with K ATP channel openers and blockers. 21 , 22 The SUR2B possesses 3 transmembrane domains (TMD0, TMD1, and TMD2), 2 cytoplasmic nucleotide binding domains (NBD1 and NBD2), and the Walker A, Walker B, and Linker L consensus sequences. 23 It functions as a regulatory subunit to mediate gating of the Kir6.1 pore by sulfonylurea drugs, such as Gli.…”

Section: Discussionmentioning

confidence: 99%

Altered KATP Channel Subunits Expression and Vascular Reactivity in Spontaneously Hypertensive Rats With Age

Liu

Duan

et al. 2016

Journal of Cardiovascular Pharmacology

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Abstract:ATP-sensitive potassium (KATP) channels link membrane excitability to metabolic state to regulate a series of biological activities including the vascular tone. However, their ability to influence hypertension is controversial. Here we aim to investigate possible alteration of KATP channel in vascular smooth muscles (VSMs) during hypertension development process. In this study, we used 16-week-old spontaneously hypertensive rats (SHRs), 49-week-old SHRs, and their age-matched Wistar-Kyoto rats to study the expression of VSM KATP subunits at the mRNA and protein level and the function of VSM KATP by observing the relaxation reactivity of isolated aorta rings to KATP modulators. We found that the expression of VSM KATP subunits Kir6.1 and sulfonylurea receptor (SUR2B) decreased during hypertension. Moreover, the expression of SUR2B and Kir6.1 in 49-week-old SHRs decreased much more than that in 16-week-old SHRs. Furthermore, the aorta rings of 49-week-old SHRs showed lower reactivity to diazoxide than 16-week-old SHRs. This study suggests that KATP channels in VSM subunits Kir6.1 and SUR2B contribute to modify the functionality of this channel in hypertension with age.

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“…These results may explain how K ATP channel activity might respond rapidly to changes in the cardiac workload in a healthy heart and in a pathological state . In addition, phosphorylating NBD1 of SUR2B also exposes the MgATP binding site, thus disrupting the interaction of the NBD core with the N‐terminal tail and, in turn, activating K ATP channels …”

Section: Cardiac Diseasesmentioning

confidence: 99%

“…23 In addition, phosphorylating NBD1 of SUR2B also exposes the MgATP binding site, thus disrupting the interaction of the NBD core with the N-terminal tail and, in turn, activating K ATP channels. 24…”

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confidence: 99%

Functional protection against cardiac diseases depends on ATP‐sensitive potassium channels

Zhu

et al. 2018

J Cellular Molecular Medi

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ATP‐sensitive potassium channels (KATP) channels are widely distributed in various tissues, including pancreatic beta cells, muscle tissue and brain tissue. KATP channels play an important role in cardioprotection in physiological/pathological situations. KATP channels are inhibited by an increase in the intracellular ATP concentration and are stimulated by an increase in the intracellular MgADP concentration. Activation of KATP channels decreases ischaemia/reperfusion injury, protects cardiomyocytes from heart failure, and reduces the occurrence of arrhythmias. KATP channels are involved in various signalling pathways, and their participation in protective processes is regulated by endogenous signalling molecules, such as nitric oxide and hydrogen sulphide. KATP channels may act as a new drug target to fight against cardiovascular disease in the development of related drugs in the future. This review highlights the potential mechanisms correlated with the protective role of KATP channels and their therapeutic value in cardiovascular diseases.

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Phosphorylation-dependent Changes in Nucleotide Binding, Conformation, and Dynamics of the First Nucleotide Binding Domain (NBD1) of the Sulfonylurea Receptor 2B (SUR2B)

Abstract: Background: Phosphorylation of SUR2B NBD1 activates ATP-sensitive K ϩ (K ATP ) channels.

Cited by 14 publications

References 97 publications

HK97 gp74 Possesses an α-Helical Insertion in the ββα Fold That Affects Its Metal Binding, cos Site Digestion, and In Vivo Activities

HK97 gp74 Possesses an α-Helical Insertion in the ββα Fold That Affects Its Metal Binding, cos Site Digestion, and In Vivo Activities

Altered KATP Channel Subunits Expression and Vascular Reactivity in Spontaneously Hypertensive Rats With Age

Functional protection against cardiac diseases depends on ATP‐sensitive potassium channels

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