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Dynamic Measurement of intracellular ph Based on Bioluminescent Bacteria

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Chemical Research in Chinese Universities 2024年第2期40卷 287-292页

作者： LI Yaohua WANG Wei State Key Laboratory of Analytical Chemistry for Life Science School of Chemistry and Chemical EngineeringChemBIC(Chemistry and Biomedicine Innovation Center)Nanjing UniversityNanjing210023P.R.China

intracellular ph(phi)is a fundamental indicator of cellular physiological state,regulating cellular state and function,and has important research *** various probes for measuring intracellular ph were available,it is challenging to reflect phi in real-time and reversible ***,we developed a whole-cell bioluminescent(BL)probe based on wild type BL bacteria,photobacterium phosphoreum(***),to determine and image *** dependence of BL intensity of *** on ph values of culture solutions was *** was found that BL intensity could respond to the change of ph values rapidly and *** further revealed that *** maintained ph homeostasis in the extracellular ph(phe)within the range of 5.0–7.0,while intracellular ph homeostasis was destroyed at the alkaline *** method opens up the enormous potential of BL bacteria as an alternative to fluorescence for monitoring and imaging phi.

关键词： Bioluminescence P.phosphoreum intracellular ph Cell imaging

Monitoring intracellular ph fluctuation with an excited-state intramolecular proton transfer-based ratiometric fluorescent sensor

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Chinese Chemical Letters 2021年第10期32卷 3057-3060页

作者： Bin Feng Yingli Zhu Jiaxin Wu Xueyan Huang Rong Song Liu Huang Xueping Feng Wenbin Zeng Xiangya School of pharmaceutical Sciences Central South UniversityChangsha 410013China The Molecular Imaging Research Center Central South UniversityChangsha 410013China Hunan Key Laboratory of Diagnostic and Therapeutic Drug Research for Chronic Diseases Central South UniversityChangsha 410013China Xiangya Hospital Central South UniversityChangsha 410013China

intracellular ph is a key parameter related to various biological and pathological *** this study,a ratiometric ph fluorescent sensor ABTT was developed harnessing the amino-type excited-state intramolecular proton transfer(ESIPT) *** on whether the ESIPT proceeds normally or not,ABTT exhibited the yellow fluorescence in acidic media,or cyan fluorescence in basic *** to the variation,ABTT behaved as a promising sensor which possessed fast and reversible response to ph change without interference from the biological substances,and exported a steady ratiometric signal(I_(478)/I_(546)).Moreover,due to the ESIPT effect,large Stokes shift and high quantum yield were also exhibited in ***,ABTT was applied for monitoring the ph changes in living cells and visualizing the ph fluctuations under oxidative stress *** results elucidated great potential of ABTT in understanding ph-dependent physiological and pathological processes.

关键词： Excited-state intramolecular proton transfer intracellular ph Sulfonamide Ratiometric sensor Fluorescent imaging

LipR functions as an intracellular ph regulator in Bacillus thuringiensis under glucose conditions

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mLife 2023年第1期2卷 58-72页

作者： Xia Cai Jiaxin Qin Xuelian Li Taoxiong Yuan Bing Yan Jun Cai Department of Microbiology College of Life SciencesNankai UniversityTianjinChina School of Life Science and Engineering Lanzhou University of TechnologyLanzhouChina Key Laboratory of Molecular Microbiology and Technology Ministry of EducationTianjinChina Tianjin Key Laboratory of Microbial Functional Genomics TianjinChina

intracellular ph critically affects various biological processes,and an appropriate cytoplasmic ph is essential for ensuring bacterial *** is the preferred carbon source for most heterotrophs;however,excess glucose often causes the accumulation of acidic metabolites,lowering the intracellular ph and inhibiting bacterial *** thuringiensis can effectively cope with glucose-induced stress;unfortunately,little is known about the regulators involved in this ***,we document that the target of the dual-function sRNA YhfH,the lipR gene,encodes a LacI-family transcription factor LipR as an intracellular ph regulator when *** BMB171 is suddenly exposed to *** glucose conditions,lipR deletion leads to early growth arrest by causing a rapid decrease in intracellular ph(~5.4).Then,the direct targets and a binding motif(GAWAWCRWTWTCAT)of LipR were identified based on the electrophoretic mobility shift assay,the DNase-I footprinting assay,and RNA sequencing,and the gapN gene encoding a key enzyme in glycolysis was directly inhibited by ***,Ni^(2+)is considered a possible effector for *** addition to YhfH,the lipR expression was coregulated by itself,CcpA,and *** study reveals that LipR plays a balancing role between glucose metabolism and intracellular ph in *** subjected to glucose stress.

关键词： glucose intracellular ph LacI‐type transcription factor LipR

Fenton reactions drive nucleotide and ATP syntheses in cancer

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Journal of Molecular Cell Biology 2018年第5期10卷 448-459页

作者： Huiyan Sun Chi Zhang Sha Cao Tao Sheng Ning Dong Ying Xu College of Computer Science and Technology Jilin University Changchun China Computational Systems Biology Lab Department of Biochemistry and Molecular Biology and institute of Bioinformatics University of Georgia GA USA School of Public Health Jilin University Changchun China The First Hospital of Jilin University Changchun China

We present a computational study of tissue transcriptomic data of 14 cancer types to address： what may drive cancer cell division？ Our analyses point to that persistent disruption of the intraceUular ph by Fenton reactions may be at the root of cancer development. Specifically, we have statistically demonstrated that Fenton reactions take place in cancer cytosoi and mitochondria across all the 14 cancer types, based on cancer tissue gene-expression data integrated via the Michaelis-Menten equation. In addition, we have shown that （i） Fenton reactions in cytosol of the disease cells will continuously increase their ph, to which the cells respond by generating net protons to keep the ph stable through a combination of synthesizing glycolytic ATPs and consuming them by nucleotide syntheses, which may drive cell division to rid of the continuously synthesized nucleotides; and （ii） Fenton reactions in mitochondria give rise to novel ways for ATP synthesis with electrons ultimately coming from H2O2, largely originated from immune cells. A model is developed to link these to cancer development, where some mutations may be selected to facilitate cell division at rates dictated by Fenton reactions.

关键词： cancer driver Fenton reaction gene-expression data intracellular ph Warburg effect

Nucleoporin 35 regulates cardiomyocyte ph homeostasis by controlling Na^(+)-H^(+)exchanger-1 expression

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Journal of Molecular Cell Biology 2015年第5期7卷 476-485页

作者： Liang Xu Lei Pan Jun Li Bijun Huang Jing Feng Changming Li ShiyiWang Erlinda The Yuan Liu Tianyou Yuan Lixiao Zhen Dandan Liang Yi Liu Li Li Yingyu Cui Xiaoyan Jiang Luying Peng Yi-Han Chen Key Laboratory of Arrhythmias of the Ministry of Education of China Shanghai East HospitalTongji University School of MedicineShanghai 200120China Research Center for Translational Medicine Shanghai East HospitalTongji University School of MedicineShanghai 200120China Institute of Medical Genetics Tongji UniversityShanghai 200092China Department of Cardiology Shanghai East HospitalTongji University School of MedicineShanghai 200120China Department of Pathology and Pathophysiology Tongji University School of MedicineShanghai 200092China

The mammalian nuclear pore complex is comprised of∼30 different nucleoporins(Nups).It governs the nuclear import of gene expression modulators and the export of *** cardiomyocytes,Na1-H1 exchanger-1(NHE1)is an integral membrane protein that exclusively regulates intracellular ph(phi)by exchanging one intracellular H1 for one extracellular ***,the role of Nups in cardiac NHE1 expression remains *** herein report that Nup35 regulates cardiomyocyte NHE1 expression by controlling the nucleo-cytoplasmic trafficking of nhe1 *** N-terminal domain of Nup35 determines nhe1 mRNA nuclear export by targeting the 5′-UTR(2412 to2213 nt)of ***35 ablationweakensthe resistance of cardiomyocytes to an acid challenge by depressingNHE1 ***,we identify thatNup35 andNHE1 are simultaneously downregulated in ischemic cardiomyocytes both in vivo and in *** expression of Nup35 effectively counteracts the anoxia-induced intracellular *** conclude that Nup35 selectively regulates cardiomyocyte phi homeostasis by posttranscriptionally controlling NHE1 *** finding reveals a novel regulatory mechanism of cardiomyocyte phi,and may provide insight into the therapeutic strategy for ischemic cardiac diseases.

关键词： cardiomyocyte intracellular ph Na^(+)-H^(+)exchanger nuclear pore complex nucleo-cytoplasmic transport

Characterization of H^+ and HCO_3^- transporters in CFPAC-1 human pancreatic duct cells

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World Journal of Gastroenterology 2006年第6期12卷 885-895页

作者： ZoltánRakonczayJr AmyFearn PéterHegy MichaelAGray BarryEArgent PéterHegyi Imre Boros institute for Cell anti Molecular Biosciences.University of Newcastle upon Tyne Medical SchoolNewcastle upon Tyne NE2 4HHUnited Kingdom First Department of Medicine Faculty of MedicineUniversity of SzegedH-6720 Szeged institute for Cell anti Molecular Biosciences.University of Newcastle upon Tyne Medical SchoolNewcastle upon Tyne NE2 4HH Hungarian Academy of Sciences Biological Research CenterInstitute of BiochemistryH-6726 Szeged

AIM： To characterize H^＋ and HCO3^- transporters in polarized CFPAC-1 human pancreatic duct cells, which were derived from a cystic fibrosis patient with the AF508 CFTR mutation. METHODS： CFPAC-1 cells were seeded at high density onto permeable supports and grown to confluence. The cells were loaded with the ph-sensitive fluorescent dye BCECF, and mounted into a perfusion chamber, which allowed the simultaneous perfusion of the basolateral and apical membranes. Transmembrane base flux was calculated from the changes in intracellular ph and the buffering capacity of the cells. RESULTS： Our results showed differential permeability to HCO3^＋/CO2 at the apical and basolateral membranes of CFPAC-1 cells. Na^＋/HCO3^- co-transporters （NBCs） and Cl^-/HCO3^- exchangers （AEs） were present on the basolateral membrane, and Na＋/H＋ exchangers （NHEs） on both the apical and basolateral membranes of the cells. Basolateral HCO3 uptake was sensitive to variations of extracellular K^＋ concentration, the membrane permeable carbonic anhydrase （CA） inhibitors acetazolamide （100 μmol/L） and ethoxyzolamide （100 μmol/L）, and was partially inhibited by H2-DIDS （600 μmol/L）. The membrane-impermeable CA inhibitor 1-N-（4-sulfamoylphenylethyl）-2,4,6-trimethylpyridine perchlorate did not have any effect on HCO3^- *** basolateral AE had a much higher activity than that in the apical membrane, whereas there was no such difference with the NHE under resting conditions. Also, 10 μmol/L forskolin did not significantly influence Cl^-/HCO3^- exchange on the apical and basolateral membranes. The administration of 250 μmol/L H2-DIDS significantly inhibited the basolateral AE. Amiloride （300 μmol/L） completely inhibited NHEs on both membranes of the cells. RT-PCR revealed the expression of pNBC1, AE2, and NHE1 mRNA. CONCLUSION： These data suggest that apart from the lack of CFTR and apical Cl^-/HCO3^- exchanger activity, CFPAC-1 cells express similar H^＋ and HCO3^- transporters to those observe

关键词： CFPAC-1 Pancreatic duct cells HCO3^- intracellular ph

Cellular physiological approach for treatment of gastric cancer

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World Journal of Gastroenterology 2014年第33期20卷 11560-11566页

作者： Atsushi Shiozaki Daisuke Ichikawa Eigo Otsuji Yoshinori Marunaka Division of Digestive Surgery Department of Surgery Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Departments of Molecular Cell physiology and Bio-Ionomics Graduate School of Medical Science Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Japan Institute for Food Education and Health St. Agnes’ University Kyoto 602-8013 Japan

Recent studies show that ion channels/transporters play important roles in fundamental cellular functions that would be involved in the cancer process. We review the evidence for their expression and functioning in human gastric cancer (GC), and evaluate the potential of cellular physiological approach in clinical management. Various types of ion channels, such as voltage-gated K+ channels, intracellular Cl- channels and transient receptor potential channels have been found to express in GC cells and tissues, and to control cell cycles. With regard to water channels, aquaporin 3 and 5 play an important role in the progression of GC. Regulators of intracellular ph, such as anion exchanger, sodium-hydrogen exchanger, vacuolar H+-ATPases and carbonic anhydrases are also involved in tumorigenesis of GC. Their pharmacological manipulation and gene silencing affect cellular behaviours, suggesting their potential as therapeutic targets for GC. Our studies indicate the intracellular Cl- concentration could act as a mediator of cellular signaling and control cell cycle progression in GC cells. Further, we demonstrate the cytocidal effects of hypotonic shock on GC cells, and indicate that the blockade of Cl- channels/transporters enhances these effects by inhibiting regulatory volume decrease. A deeper understanding of molecular mechanisms may lead to the discovery of these cellular physiological approaches as a novel therapeutic strategy for GC.

关键词： Gastric cancer Ion channels Water chan-nels intracellular ph intracellular chloride Osmolality

A New Ratio-Metric ph Probe,“ThiAKS Green”for Live-Cell ph Measurements

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photonic Sensors 2023年第1期13卷 66-75页

作者： Ali AKYOL Doruk BAYKAL Akin AKDAG Özge SENSOY Çagdas Devrim SON Middle East Technical University Department of Biological SciencesAnkara 06800Turkey Middle East Technical University Department of ChemistryAnkara 06800Turkey Istanbul Medipol University School of Engineering and Natural Sciencesistanbul 34810Turkey

Deviation of the H+concentration from optimum values within the organelles is closely associated with irregular cellular functions that cause the onset of various ***,determining subcellular ph values in live cells and tissues is valuable for diagnostic *** this study,we report a novel ratiometric fluorescence probe 1H-pyrazole-3-carboxylic acid,4-(benzo[d]thiazol-2-yl)-3-(2,4-dihydroxy-3-methylphenyl)-1H-pyrazole-5-carboxylicacid4-(2-benz othiazolyl)-5-(2,4-dihydroxy-3-methylphenyl),to which we will refer as ThiAKS Green(Thiazole AKyol shifting green),that is ph *** results presented here show that the probe can penetrate the celi membrane in iess than 30minutes and does not show any detectable *** measured color shifts up on ph change are linear and most significant around physiological ph(pKa=7.45),thus making this probe suitable for live-cell imaging and intracellular ph *** the long-incubation periods following the application of the probe and the fluorescent microscopy measurements,it shows stable properties and is easy to detect in live *** conclusion,the results suggest that ThiAKS Green can be used to obtain precise information on the H+distribution at various compartments of the live cells.

关键词： Fluorescent probe ratiometric sensor intracellular ph cell imaging fluorescent microscopy