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Ca2+ single

Antonio Villalobo, María González-Muñoz, Martin W Berchtold
The appearance of modular proteins is a widespread phenomenon during the evolution of proteins. The combinatorial arrangement of different functional and/or structural domains within a single polypeptide chain yields a wide variety of activities and regulatory properties to the modular proteins. In this review, we will discuss proteins, that in addition to their catalytic, transport, structure, localization or adaptor functions, also have segments resembling the helix-loop-helix EF-hand motifs found in Ca2+ -binding proteins, such as calmodulin (CaM)...
March 15, 2019: Cellular and Molecular Life Sciences: CMLS
Kristen M Varney, Paul T Wilder, Raquel Godoy-Ruiz, Filippo Mancia, David J Weber
Vitamin A is a necessary nutrient for all mammals, and it is required for the transcription of many genes and vital for vision. While fasting, the vitamin A alcohol form (Retinol) from storage in the liver is mobilized and transported through the bloodstream while bound to retinol binding protein (RBP). Details of how exactly vitamin A is released from RBP and taken into the cells are still unclear. As part of the effort to elucidate the specifics of this process, single-particle cryo-electron microscopy structural studies of STRA6 (the RBP receptor 75-kDa transmembrane receptor protein) were recently reported by Chen et al...
March 14, 2019: Biomolecular NMR Assignments
Leelatulasi Salwoom, Raja Noor Zaliha Raja Abd Rahman, Abu Bakar Salleh, Fairolniza Mohd Shariff, Peter Convey, Mohd Shukuri Mohamad Ali
In recent years, studies on psychrophilic lipases have become an emerging area of research in the field of enzymology. The study described here focuses on the cold-adapted organic solvent tolerant lipase strain Pseudomonas sp. LSK25 isolated from Signy Station, South Orkney Islands, maritime Antarctic. Strain LSK25 lipase was successfully cloned, sequenced, and over-expressed in an Escherichia coli system. Sequence analysis revealed that the lipase gene of Pseudomonas sp. LSK25 consists of 1432 bp, lacks an N-terminal signal peptide and encodes a mature protein consisting of 476 amino acids...
March 13, 2019: International Journal of Molecular Sciences
Marie Mita, Motoki Ito, Kazuki Harada, Izumi Sugawara, Hiroshi Ueda, Takashi Tsuboi, Tetsuya Kitaguchi
Glucose is the most important energy source for living animals. Here, we developed a series of single fluorescent protein (FP)-based glucose indicators, named as "Green Glifons", to understand the hierarchal and mutual relationships between molecules involved in energy metabolism. Three indicators showed a different EC50 for glucose (50 µM, 600 µM and 4,000 µM), producing a ~7-fold change in fluorescence intensity in response to glucose. The indicators could visualize glucose dynamics in the cytoplasm, plasma membrane, nucleus and mitochondria of living HeLa cells and in vivo, in the pharyngeal muscle of C...
March 14, 2019: Analytical Chemistry
Yoshitake Cho, Shizuko Tachibana, Bethany C Hazen, James J Moresco, John R Yates, Bernard Kok, Enrique Saez, Robert S Ross, Aaron P Russell, Anastasia Kralli
OBJECTIVE: Endurance exercise training remodels skeletal muscle, leading to increased mitochondrial content and oxidative capacity. How exercise entrains skeletal muscle signaling pathways to induce adaptive responses remains unclear. In past studies, we identified Perm1 (PGC-1 and ERR induced regulator, muscle 1) as an exercise-induced gene and showed that Perm1 overexpression elicits similar muscle adaptations as endurance exercise training. The mechanism of action and the role of Perm1 in exercise-induced responses are not known...
February 27, 2019: Molecular Metabolism
Kyaw Thiha, Yoichi Mashimo, Hiroyuki Suzuki, Hiromichi Hamada, Akira Hata, Toshiro Hara, Toshihiro Tanaka, Kaoru Ito, Yoshihiro Onouchi
ORAI1 encodes a calcium channel essential in the store-operated calcium entry mechanism. A previous genetic association study identified a rare in-frame insertion variant of ORAI1 conferring Kawasaki disease (KD). To deepen our understanding of the involvement of rare variants of ORAI1 in KD pathogenesis, we investigated 3812 patients with KD and 2644 healthy individuals for variations in the protein-coding region of ORAI1. By re-sequencing the study participants' DNA, 27 variants with minor allele frequencies (MAFs) < 0...
March 11, 2019: Journal of Human Genetics
Aleksandra Klimas, Gloria Ortiz, Steven C Boggess, Evan W Miller, Emilia Entcheva
Combined optogenetic stimulation and optical imaging permit scalable, contact-free high-throughput probing of cellular electrophysiology and optimization of stem-cell derived excitable cells, such as neurons and muscle cells. We report a new "on-axis" configuration (combined single optical path for stimulation and for multiparameter imaging) of OptoDyCE, our all-optical platform for studying human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) and other cell types, optically driven by Channelrhodopsin2 (ChR2)...
March 5, 2019: Progress in Biophysics and Molecular Biology
Dongqing Fang, Wei Huang, Dagmara S Antkiewicz, Yuqin Wang, Reza Bashiri Khuzestani, Yang Zhang, Jing Shang, Martin M Shafer, Lingyan He, James J Schauer, Yuanxun Zhang, Shuo Zhao
Size-resolved particulate matter (PM) was collected at the Heshan Super-Station in the Pearl River Delta (PRD) region, China, to evaluate their chemical characteristics and potential health risks. The chemical mass closures illustrate that the dominant fraction in coarse (2.5 μm < Dp  < 10 μm) PM was dust, while organic matter made up a substantial portion of both fine (0.1 < Dp  < 2.5 μm) and ultra-fine (Dp  < 0.10 μm) PM fractions. The size distribution of most PM components varied substantially...
March 7, 2019: Environmental Science and Pollution Research International
Robert John Holash, Brian R MacIntosh
Skeletal muscle contraction is initiated when an action potential triggers the release of Ca2+ into the sarcomere in a process referred to as excitation-contraction coupling. The speed and scale of this process makes direct observation very challenging and invasive. To determine how the concentration of Ca2+ changes within the myofibril during a single activation, several simulation models have been developed. These models follow a common pattern; divide the half sarcomere into a series of compartments, then use ordinary differential equations to solve reactions occurring within and between the compartments...
March 7, 2019: PLoS Computational Biology
Jingxiu Bi, Fan Feng, Jia Li, Jinjuan Mao, Mingxiao Ning, Xiaowen Song, Jia Xie, Jing Tang, Bin Li
C-type lectins are one of the pattern-recognition proteins involved in innate immunity in invertebrates. Although there are 16 C-type lectin genes that have been identified in the genome of Tribolium castaneum, their functions and mechanisms in innate immunity remain unknown. Here, we identified one C-type lectin ortholog, TcCTL6 (TC003708), by sequencing random clones from the cDNA library of the coleopteran beetle, T. castaneum. TcCTL6 contains a 654 bp open reading frame encoding a protein of 217 amino acids that includes a single carbohydrate-recognition domain (CRD)...
March 7, 2019: Insect Molecular Biology
Gizem Keceli, Ananya Majumdar, Chevon N Thorpe, Seungho Jun, Carlo G Tocchetti, Dong I Lee, James E Mahaney, Nazareno Paolocci, John P Toscano
Nitroxyl (HNO) positively modulates myocardial function by accelerating Ca2+ reuptake into the sarcoplasmic reticulum (SR). HNO-induced enhancement of myocardial Ca2+ cycling and function is due to the modification of cysteines in the transmembrane domain of phospholamban (PLN), which results in activation of SR Ca2+ -ATPase (SERCA2a) by functionally uncoupling PLN from SERCA2a. However, which cysteines are modified by HNO, and whether HNO induces reversible disulfides or single cysteine sulfinamides (RS(O)NH2 ) that are less easily reversed by reductants, remain to be determined...
March 6, 2019: Journal of General Physiology
Michitaka Amioka, Yukiko Nakano, Hidenori Ochi, Yuko Onohara, Akinori Sairaku, Takehito Tokuyama, Chikaaki Motoda, Hiroya Matsumura, Shunsuke Tomomori, Naoya Hironobe, Yousaku Okubo, Sho Okamura, Kazuaki Chayama, Yasuki Kihara
BACKGROUND: Atrial fibrillation (AF) recurrence after radiofrequency catheter ablation (RFCA) still remains a serious issue. Ca2+ handling has a considerable effect on AF recurrence. The histidine-rich calcium-binding protein (HRC) genetic single nucleotide polymorphism (SNP), rs3745297 (T>G, Ser96Ala), is known to cause a sarcoplasmic reticulum Ca2+ leak. We investigated the association between HRC Ser96Ala and AF recurrence after RFCA in paroxysmal AF (PAF) patients. METHODS AND RESULTS: We enrolled PAF patients who underwent RFCA (N = 334 for screening and N = 245 for replication) and were genotyped for HRC SNP (rs3745297)...
2019: PloS One
Liang Zhang, Kaouter Bouadjel, Boris Manoury, Grégoire Vandecasteele, Rodolphe Fischmeister, Véronique Leblais
BACKGROUND AND PURPOSE: PDE upregulation is associated with several vascular diseases and better understanding of the respective role of each PDE family in controlling subcellular pools of cyclic nucleotides in vascular cells is needed. We investigated the respective role of PDE1, PDE5 and PDE9 in controlling intracellular cAMP and/or cGMP concentrations ([cAMP]i , [cGMP]i ) in cultured rat aortic smooth muscle cells (RASMCs). EXPERIMENTAL APPROACH: By using a pharmacological approach with selective PDE1, PDE5 and PDE9 inhibitors (PF-04471141, sildenafil and PF-04447943, respectively), cAMP- and cGMP-PDE activities were measured by a radioenzymatic assay in protein extracts...
March 1, 2019: British Journal of Pharmacology
Wenting Sui, Kunyuan Guo, Li Li, Shenkui Liu, Tetsuo Takano, Xinxin Zhang
Eukaryotic nucleases are involved in processes such as DNA restriction digestion, repair, recombination, transposition, and programmed cell death (PCD). Studies on the role of nucleases have mostly focused on PCD during plant development, while the information on nucleases involved in responses to different abiotic stress conditions remains limited. Here, we identified a Ca2+ -dependent nuclease, AtCaN2, in Arabidopsis thaliana and characterized its activity, expression patterns, and involvement in plant responses to salt stress...
April 2019: Plant Science: An International Journal of Experimental Plant Biology
Elisa Greotti, Paola Capitanio, Andrea Wong, Tullio Pozzan, Paola Pizzo, Diana Pendin
An imbalance in Ca2+ homeostasis represents an early event in the pathogenesis of Alzheimer's disease (AD). Presenilin-1 and -2 (PS1 and PS2) mutations, the major cause of familial AD (FAD), have been extensively associated with alterations in different Ca2+ signaling pathways, in particular those handled by storage compartments. However, FAD-PSs effect on organelles Ca2+ content is still debated and the mechanism of action of mutant proteins is unclear. To fulfil the need of a direct investigation of intracellular stores Ca2+ dynamics, we here present a detailed and quantitative single-cell analysis of FAD-PSs effects on organelle Ca2+ handling using specifically targeted, FRET (Fluorescence/Förster Resonance Energy Transfer)-based Ca2+ indicators...
February 23, 2019: Cell Calcium
Michele Cianci, Max Nanao, Thomas R Schneider
Harnessing the anomalous signal from macromolecular crystals with volumes of less than 10 000 µm3 for native phasing requires careful experimental planning. The type of anomalous scatterers that are naturally present in the sample, such as sulfur, phosphorus and calcium, will dictate the beam energy required and determine the level of radiation sensitivity, while the crystal size will dictate the beam size and the sample-mounting technique, in turn indicating the specifications of a suitable beamline. On the EMBL beamline P13 at PETRA III, Mesh&Collect data collection from concanavalin A microcrystals with linear dimensions of ∼20 µm or less using an accordingly sized microbeam at a wavelength of 1...
February 1, 2019: Acta Crystallographica. Section D, Structural Biology
Zsuzsanna É Magyar, Gyula Diszházi, Judit Péli-Szabó, Péter Szentesi, Claude Collet, László Csernoch, Péter Nánás, János Almássy
Very recently, the diamide insecticide chlorantraniliprole was shown to induce Ca2+-release from sarcoplasmic reticulum (SR) vesicles isolated from mammalian skeletal muscle through the activation of the SR Ca2+ channel ryanodine receptor. As this result raises severe concerns about the safety of this chemical, we aimed to learn more about its action. To this end, single-channel analysis was performed, which showed that chlorantraniliprole induced high-activity bursts of channel opening that accounts for the Ca2+-releasing action described before...
March 1, 2019: General Physiology and Biophysics
Wei-Guang Ding, Ayami Tano, Xinya Mi, Akiko Kojima, Tomoyoshi Seto, Hiroshi Matsuura
BACKGROUND/AIMS: The phenylalkylamine class of L-type Ca2+ channel antagonist verapamil prolongs the effective refractory period (ERP) of human atrium, which appears to contribute to the efficacy of verapamil in preventing reentrant-based atrial arrhythmias including atrial fibrillation. This study was designed to investigate the molecular and electrophysiological mechanism underlying the action of verapamil on human Kv1.5 (hKv1.5) channel that determines action potential duration and ERP in human atrium...
2019: Cellular Physiology and Biochemistry
Ashekul Islam, Talha Bin Emran, Daisuke S Yamamoto, Mitsuhiro Iyori, Fitri Amelia, Yenni Yusuf, Ririka Yamaguchi, Md Shah Alam, Henrique Silveira, Shigeto Yoshida
The saliva of hematophagous arthropods is enriched with a complex mixture of antihemostatic molecules, the biological functions of which are largely unknown. Anopheline antiplatelet protein (AAPP) from malaria vector mosquito exhibits strong antiplatelet activity when bound directly to host collagen by its C-terminus and through its N-terminus with Ca2+ -binding activity. To investigate the biological functions of AAPP in blood feeding behavior and malaria transmission, we generated transgenic Anopheles stephensi mosquito lines expressing anti-AAPP antibody single-chain fragment (scFv) in their salivary glands...
February 28, 2019: Scientific Reports
Bradley Quade, Marcial Camacho, Xiaowei Zhao, Marta Orlando, Thorsten Trimbuch, Junjie Xu, Wei Li, Daniela Nicastro, Christian Rosenmund, Josep Rizo
Munc13-1 plays a crucial role in neurotransmitter release. We recently proposed that the C-terminal region encompassing the C1 , C2 B, MUN and C2 C domains of Munc13-1 (C1 C2 BMUNC2 C) bridges the synaptic vesicle and plasma membranes through interactions involving the C2 C domain and the C1 -C2 B region. However, the physiological relevance of this model has not been demonstrated. Here we show that C1 C2 BMUNC2 C bridges membranes through opposite ends of its elongated structure. Mutations in putative membrane-binding sites of the C2 C domain disrupt the ability of C1 C2 BMUNC2 C to bridge liposomes and to mediate liposome fusion in vitro...
February 28, 2019: ELife
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