Characterization of endogenous Kv1.3 channel isoforms in T cells. DOI Creative Commons
Julia Serna

Опубликована: Янв. 1, 2023

The voltage-gated potassium channel Kv1.3 plays a crucial role in T-cell activation and is considered promising target for the treatment of autoimmune diseases.However, lack reliable antibodies has prevented its accurate detection study under endogenous conditions, so that most published studies have been conducted heterologous systems.To address this limitation, we engineered Jurkat line expressing channels tagged with signal peptide to investigate expression localization native channels, their associated T cell physiological responses.Using CRISPR-Cas9 tool, inserted Flag-Myc at C terminus KCNA3 gene.Basal activated were assessed through western blot analysis imaging techniques.Surprisingly, besides canonical (54 KDa), identified two additional isoforms distinct N termini: longer isoform (70 KDa) truncated (43 KDa).All three showed upregulation after activation.Our focus was on characterizing (short form, SF) as it had not previously described could be present available Kv1.3-/-mouse models.Overexpressing SF HEK cells generated Kv1.3-like currents smaller amplitudes, which, unlike Kv1.3, did induce proliferation.To explore system, both knockout clone only isoform.While localized primarily plasma membrane, remained intracellular, accumulating perinuclearly.Consequently, lacked currents, exhibited depolarized resting membrane potential (EM), reduced Ca 2+ influx, diminished increases intracellular calcium ([Ca ]i) upon stimulation.Functional characterization these revealed differential contributions signaling pathways involved immunological synapse formation.In conclusion, alternative translation initiation generates least functional roles.Importantly, some functions do require formation by proteins.

Язык: Английский

Structure of an open KATP channel reveals tandem PIP2 binding sites mediating the Kir6.2 and SUR1 regulatory interface DOI Creative Commons
Camden Driggers, Yi‐Ying Kuo, Phillip Zhu

и другие.

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Март 20, 2024

Abstract ATP-sensitive potassium (K ATP ) channels, composed of four pore-lining Kir6.2 subunits and regulatory sulfonylurea receptor 1 (SUR1) subunits, control insulin secretion in pancreatic β-cells. K channel opening is stimulated by PIP 2 inhibited ATP. Mutations that increase reduce inhibition cause neonatal diabetes. Although considerable evidence has implicated a role for function, previously solved open-channel structures have lacked bound , mechanisms which regulates channels remain unresolved. Here, we report the cryoEM structure harboring diabetes mutation Kir6.2-Q52R, open conformation, to amphipathic molecules consistent with natural C18:0/C20:4 long-chain PI(4,5)P at two adjacent binding sites between SUR1 Kir6.2. The canonical site conserved among -gated Kir channels. non-canonical forms interface SUR1. Functional studies demonstrate both determine activity. pore associated twist cytoplasmic domain rotation N-terminal transmembrane SUR1, widens inhibitory pocket disfavor binding. conformation particularly stabilized Kir6.2-Q52R residue through cation-π bonding SUR1-W51. Together, these results uncover cooperation gating, explain antagonistic regulation ATP, provide putative mechanism stabilizes an

Язык: Английский

Процитировано

14
KATP channel mutations in congenital hyperinsulinism: Progress and challenges towards mechanism-based therapies DOI Creative Commons
Assmaa ElSheikh, Show‐Ling Shyng

Frontiers in Endocrinology, Год журнала: 2023, Номер 14

Опубликована: Март 28, 2023

Congenital hyperinsulinism (CHI) is the most common cause of persistent hypoglycemia in infancy/childhood and a serious condition associated with severe recurrent attacks due to dysregulated insulin secretion. Timely diagnosis effective treatment are crucial prevent that may lead life-long neurological complications. In pancreatic β-cells, adenosine triphosphate (ATP)-sensitive K

Язык: Английский

Процитировано

17
Dynamic duo: Kir6 and SUR in K ATP channel structure and function DOI Creative Commons

Bruce L. Patton,

Phillip Zhu, Assmaa ElSheikh

и другие.

Channels, Год журнала: 2024, Номер 18(1)

Опубликована: Март 15, 2024

K

Язык: Английский

Процитировано

6
ATP-sensitive potassium channel opener, Nicorandil, inhibits NF-κB/AIM2/GSDMD pathway activation to protect against neuroinflammation in ischemic stroke DOI Creative Commons

Chenming Zhao,

Xiaojuan Fu,

Zhuoying Yang

и другие.

Neurochemistry International, Год журнала: 2024, Номер 179, С. 105810 - 105810

Опубликована: Июль 26, 2024

The absent in melanoma 2 (AIM2) inflammasome contributes to ischemic brain injury by inducing cell pyroptosis and inflammatory responses. Our research group has previously demonstrated that ATP-sensitive potassium channels (KATP channels) openers can modulate neuronal synaptic plasticity post-ischemic stroke for neuroprotection. However, the specific mechanisms of KATP response following remain unclear. Here, we assessed cellular damage observing changes BV-2 morphology viability. 2,3,5-Triphenyl tetrazolium chloride (TTC) staining, mNSS scoring, Nissl TdT-mediated dUTP nick end labeling (TUNEL) staining were used evaluate behavioral deficits, severity, mice subjected middle cerebral artery occlusion (MCAO). Quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA) measure nuclear factor-kappaB (NF-κB) activation vivo vitro. We observed AIM2 protein expression was upregulated localized within cytoplasm cells. Notably, low-dose Nicorandil treatment reduced cytokine secretion pyroptosis-related expression, including AIM2, cleaved cysteinyl aspartate-specific protease-1 (cleaved caspase-1), Gasdermin D N-terminal (GSDMD-NT). Further investigations revealed channel inhibitor 5-HD p-NF-κB p65, NF-κB p-IκBα reversing Nicorandil's neuroprotective effect vivo. In summary, our results suggest may serve as a potential therapeutic option stroke. Targeting represents effective strategies inhibiting neuroinflammation.

Язык: Английский

Процитировано

6
Molecular Dynamics Simulation of Kir6.2 Variants Reveals Potential Association with Diabetes Mellitus DOI Creative Commons

Mohamed E. Elangeeb,

Imadeldin Elfaki, Ali M. S. Eleragi

и другие.

Molecules, Год журнала: 2024, Номер 29(8), С. 1904 - 1904

Опубликована: Апрель 22, 2024

Diabetes mellitus (DM) represents a problem for the healthcare system worldwide. DM has very serious complications such as blindness, kidney failure, and cardiovascular disease. In addition to bad socioeconomic impacts, it influences patients their families communities. The global costs of its are huge expected rise by year 2030. is caused genetic environmental risk factors. Genetic testing will aid in early diagnosis identification susceptible individuals or populations using ATP-sensitive potassium (KATP) channels present different tissues pancreas, myocardium, myocytes, nervous tissues. respond concentrations blood sugar, stimulation hormones, ischemic conditions. pancreatic cells, they regulate secretion insulin glucagon. Mutations KCNJ11 gene that encodes Kir6.2 protein (a major constituent KATP channels) were reported be associated with Type 2 DM, neonatal diabetes (NDM), maturity-onset young (MODY). harbors binding sites ATP phosphatidylinositol 4,5-diphosphate (PIP2). inhibits channel, while (PIP2) activates it. A mutation at tyrosine330 (Y330) was demonstrated reduce inhibition predisposes NDM. this study, we examined effect mutations on structure bioinformatics tools molecular dynamic simulations (SIFT, PolyPhen, SNAP2, PANTHER, PhD&SNP, SNP&Go, I-Mutant, MuPro, MutPred, ConSurf, HOPE, GROMACS). Our results indicated M199R, R201H, R206H, Y330H influence function therefore may cause DM. We conclude MD useful techniques predict effects structure. addition, variant These require further verification protein–protein interactions, function, case-control studies.

Язык: Английский

Процитировано

5
Apigenin potentiates glucose-stimulated insulin secretion through the PKA-MEK kinase signaling pathway independent of K-ATP channels DOI Open Access

Falak Shahab,

Abdul Hameed, Akhtar Ali

и другие.

Biomedicine & Pharmacotherapy, Год журнала: 2024, Номер 177, С. 116986 - 116986

Опубликована: Июнь 20, 2024

Apigenin, a natural bioflavonoid, is reported as an anti-diabetic agent since it possesses the ability to inhibit α-glucosidase activity, cause stimulation of insulin action and secretion, manage ROS, prevent diabetes complications. Apigenin was identified new secretagogue that enhances glucose-stimulated secretion seems like better antidiabetic drug candidate. Here we explored insulinotropic mechanism(s) apigenin in vitro mice islets vivo diabetic rats.

Язык: Английский

Процитировано

4
Human ATP-binding proteins: Structural features, functional diversity, and pharmacotherapeutic potential in disease: A review DOI

Letong Li,

Shanshan Wang,

Songsen Fu

и другие.

International Journal of Biological Macromolecules, Год журнала: 2025, Номер unknown, С. 142303 - 142303

Опубликована: Март 1, 2025

Язык: Английский

Процитировано

0
Harnessing virtual screening and MD simulations: a multistage approach to identifying potent and nontoxic agonists for protein kinase A DOI

Muneeb Ali,

Nadeem Ahmad,

Madiha Sardar

и другие.

Molecular Diversity, Год журнала: 2025, Номер unknown

Опубликована: Май 26, 2025

Язык: Английский

Процитировано

0
Structure of an open KATPchannel reveals tandem PIP2binding sites mediating the Kir6.2 and SUR1 regulatory interface DOI Open Access
Camden Driggers, Yi‐Ying Kuo, Phillip Zhu

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Авг. 1, 2023

ATP-sensitive potassium (K

Процитировано

4
Molecular study of the KCNJ11 gene and its correlation with Prakriti to preventing and managing type 2 diabetes DOI Creative Commons
S Singh, Sangeeta Gehlot, Neeraj Agrawal

и другие.

Journal of Traditional and Complementary Medicine, Год журнала: 2024, Номер 14(5), С. 494 - 500

Опубликована: Янв. 11, 2024

In Ayurveda, every individual is believed to possess a unique entity known as Prakriti, which distinguishes them from others physically, physiologically, and psychologically. This also determines an individual's response particular stimulus, it that such responses are not solely determined by genetics. The present research aims validate the Ayurvedic concept of Prakriti modern molecular perspective strengthen personalized precise treatment approach. A study was conducted investigate role KCNJ11gene in susceptibility individuals type 2 diabetes mellitus (T2DM) with their metabolic status. involved allele mining on three major groups - Vata, Pitta, Kapha 112 patients T2DM healthy individuals. KCNJ11 gene, responsible for insulin secretion membrane pore formation, analyzed determine different types T2DM. MutPred tool predicted cause disease-related amino acid substitution. According study, only Pitta were diagnosed diabetes, while all control group protein model prepared, changes resulting mutations observed each sequence, both synonymous non-synonymous mutations. Ultimately, these contributed manifestation Based findings, appears may experience function due nonsynonymous differences acids at level.

Язык: Английский

Процитировано

1