Direct detection of SERCA calcium transport and small-molecule inhibition in giant unilamellar vesicles.

Abstract:

:We have developed a charge-mediated fusion method to reconstitute the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) in giant unilamellar vesicles (GUV). Intracellular Ca2+ transport by SERCA controls key processes in human cells such as proliferation, signaling, and contraction. Small-molecule effectors of SERCA are urgently needed as therapeutics for Ca2+ dysregulation in human diseases including cancer, diabetes, and heart failure. Here we report the development of a method for efficiently reconstituting SERCA in GUV, and we describe a streamlined protocol based on optimized parameters (e.g., lipid components, SERCA preparation, and activity assay requirements). ATP-dependent Ca2+ transport by SERCA in single GUV was detected directly using confocal fluorescence microscopy with the Ca2+ indicator Fluo-5F. The GUV reconstitution system was validated for functional screening of Ca2+ transport using thapsigargin (TG), a small-molecule inhibitor of SERCA currently in clinical trials as a prostate cancer prodrug. The GUV system overcomes the problem of inhibitory Ca2+ accumulation for SERCA in native and reconstituted small unilamellar vesicles (SUV). We propose that charge-mediated fusion provides a widely-applicable method for GUV reconstitution of clinically-important membrane transport proteins. We conclude that GUV reconstitution is a technological advancement for evaluating small-molecule effectors of SERCA.

authors

Bian T,Autry JM,Casemore D,Li J,Thomas DD,He G,Xing C

doi

10.1016/j.bbrc.2016.10.096

subject

Has Abstract

pub_date

2016-12-09 00:00:00

pages

206-211

issue

3-4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(16)31702-8

journal_volume

481

pub_type

杂志文章
  • Sequence of a human keratin 13 specific cDNA encompassing coil 1B through the 3' end.

    abstract::An expression library established in lambda gt11 with cDNA from squamous epithelium of the human upper digestive tract was screened with an antibody raised against keratin 13. A 1.2 kb fragment from the most strongly reacting plaque was sequenced and compared to known type I keratin sequences. The highest degree of ho...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/0006-291x(89)90847-4

    authors: Schulz P,Wachter E,Hochstrasser K,Wild AG,Mischke D

    更新日期:1989-08-15 00:00:00

  • Conformational dynamics of loops L11 and L12 of kinesin as revealed by spin-labeling EPR.

    abstract::The EPR spectra of the spin labels attached to loops L11 and L12 of kinesin were resolved into slow (rotational correlation time, tau=10-45 ns) and fast (tau=2 ns) components. The fraction of the slow component increased considerably when kinesin was complexed with a microtubule (MT). On MT binding and in the presence...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2007.10.043

    authors: Yamada MD,Maruta S,Yasuda S,Kondo K,Maeda H,Arata T

    更新日期:2007-12-21 00:00:00

  • Role of N-glycan-dependent quality control in the cell-surface expression of the AT1 receptor.

    abstract::Most G protein-coupled receptors (GPCRs) are N-glycosylated proteins but the role of this post-translational modification in GPCR biosynthesis has not been extensively studied. We previously showed that the non-glycosylated AT(1) receptor is inefficiently expressed at the cell surface. In this study, we addressed whet...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2005.12.016

    authors: Lanctôt PM,Leclerc PC,Escher E,Guillemette G,Leduc R

    更新日期:2006-02-10 00:00:00

  • Isolation of endo A cDNA from mouse 8-cell stage embryos.

    abstract::To analyse the species of genes expressed in a cleavage stage mouse embryo, we have constructed a cDNA library containing 3.0 x 10(5) independent clones from about 2 x 10(3) embryos at the 8-cell stage of development. Endo A cDNA prepared from parietal yolk sac endoderm like PYS-2 cells was used to screen the library....

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/0006-291x(88)90223-9

    authors: Nozaki M,Murata K,Morita T,Matsushiro A

    更新日期:1988-08-15 00:00:00

  • Histidine residues 102 and 117 of MelC1 play different roles in the chaperone function for Streptomyces apotyrosinase.

    abstract::MelC1 regulates the copper incorporation and secretion of Streptomyces apotyrosinase (MelC2) via a transient, competent complex formation. His-102 and His-117 of the chaperone-like MelC1 are known to play important roles in this trans-activation activity of MelC1. In this study, we studied the side chain requirement a...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1006/bbrc.1995.2307

    authors: Liaw LL,Lee YH

    更新日期:1995-09-14 00:00:00

  • Midkine is a heat and acid stable polypeptide capable of enhancing plasminogen activator activity and neurite outgrowth extension.

    abstract::We studied about the physicochemical stability of a novel heparin-binding growth/differentiation factor, midkine (MK). It was found that synthetic human MK was heat and acid stable. Neither incubation at 80 degrees C for 90 sec nor treatment at low pH affected the elution profile of MK molecule on the high performance...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1006/bbrc.1995.2661

    authors: Kojima S,Inui T,Muramatsu H,Kimura T,Sakakibara S,Muramatsu T

    更新日期:1995-11-13 00:00:00

  • Stimulus-induced phosphorylation of PKC theta at the C-terminal hydrophobic-motif in human T lymphocytes.

    abstract::Protein kinase C (PKC) is a family of serine/threonine kinases whose activity is controlled, in part, by phosphorylation on three conserved residues that are located on the catalytic domain of the enzyme, known as the activation-loop, the turn-motif, and the C-terminal hydrophobic-motif sites. Using a panel of phospho...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2005.06.136

    authors: Freeley M,Volkov Y,Kelleher D,Long A

    更新日期:2005-08-26 00:00:00

  • Identification and validation of reference genes for expression studies in a rat model of neuropathic pain.

    abstract::Neuropathic pain is triggered by damage to or as a result of the dysfunction of the somatosensory nervous system. Gene expression profiling using DNA microarray and real-time PCR have emerged as powerful tools for the elucidation of pain-specific pathways and identification of candidate biomarkers and therapeutic targ...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2010.08.106

    authors: Wan G,Yang K,Lim Q',Zhou L,He BP,Wong HK,Too HP

    更新日期:2010-10-01 00:00:00

  • Structural requirements for O2 sensing by the human tandem-P domain channel, hTREK1.

    abstract::TREK1 is a member of the tandem-P domain K+ channel family which is expressed almost exclusively in the nervous system. It is modulated by a number of important factors including arachidonic acid and cell swelling. Since both factors are associated with brain ischemia, it has been suggested that activation of TREK1 ma...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2005.04.042

    authors: Miller P,Kemp PJ,Peers C

    更新日期:2005-06-17 00:00:00

  • Sepsis-induced muscle proteolysis is prevented by a proteasome inhibitor in vivo.

    abstract::Sepsis-induced muscle proteolysis mainly reflects ubiquitin-proteasome-dependent protein degradation. The effect of in vivo administration of a proteasome inhibitor on muscle protein breakdown during sepsis is not known. We treated rats with the proteasome inhibitor N-benzyloxycarbonyl-Ile-Glu-(O-t-butyl)-Ala-leucinal...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1006/bbrc.2000.2398

    authors: Fischer D,Gang G,Pritts T,Hasselgren PO

    更新日期:2000-04-02 00:00:00

  • A computergraphic determination of the chemotactic peptide preferred conformation.

    abstract::Replacement of leucine in the chemotactic peptide For-Met-Leu-Phe by the sterically constrained amino acids alpha-aminoisobutyric acid and aminocyclohexanecarboxylic acid affords compounds of equal or greater activity than the parent. NMR studies indicate that the parent compound is present as a beta-sheet in solution...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/s0006-291x(88)80287-0

    authors: Semus SF,Becker EL,Toniolo C,Freer RJ

    更新日期:1988-12-15 00:00:00

  • Nordihydroguaiaretic acid inhibits IFN-gamma-induced STAT tyrosine phosphorylation in rat brain astrocytes.

    abstract::The Janus kinase (JAK) and signal transducers and activators of transcription (STAT) signal cascades are major pathways that mediate the inflammatory functions of interferon-gamma (IFN-gamma), an important pro-inflammatory cytokine. Therefore, regulation of JAK/STAT signaling should modulate IFN-gamma-mediated inflamm...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2005.01.025

    authors: Jeon SB,Ji KA,You HJ,Kim JH,Jou I,Joe EH

    更新日期:2005-03-11 00:00:00

  • Rescue of alpha-synuclein cytotoxicity by insulin-like growth factors.

    abstract::While mutation of alpha-synuclein is a cause of autosomal-dominant Parkinson's disease (PD), it is still elusive as to how alpha-synuclein is involved in the pathogenesis of PD. Here, we show that dopamine-dependent accumulation of alpha-synuclein in cultured cells results in apoptosis. Furthermore, activation of insu...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2009.05.089

    authors: Kao SY

    更新日期:2009-07-31 00:00:00

  • The histone demethylase PHF8 is an oncogenic protein in human non-small cell lung cancer.

    abstract::PHF8 is a JmjC domain-containing protein and erases repressive histone marks including H4K20me1 and H3K9me1/2. It binds to H3K4me3, an active histone mark usually located at transcription start sites (TSSs), through its plant homeo-domain, and is thus recruited and enriched in gene promoters. PHF8 is involved in the d...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2014.07.076

    authors: Shen Y,Pan X,Zhao H

    更新日期:2014-08-15 00:00:00

  • Autocrine galectin-1 promotes collective cell migration of squamous cell carcinoma cells through up-regulation of distinct integrins.

    abstract::We found that high galectin-1 (Gal-1) mRNA levels were associated with invasive squamous cell carcinoma (SCC) cells that expressed Snail, an epithelial-to-mesenchymal transition (EMT) regulator. Both Gal-1 overexpression and soluble Gal-1 treatment accelerated invasion and collective cell migration, along with activat...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2013.10.152

    authors: Rizqiawan A,Tobiume K,Okui G,Yamamoto K,Shigeishi H,Ono S,Shimasue H,Takechi M,Higashikawa K,Kamata N

    更新日期:2013-11-29 00:00:00

  • Porcine guanylin and uroguanylin: cDNA sequences, deduced amino acid sequences, and biological activity of the chemically synthesized peptides.

    abstract::Guanylin and uroguanylin are structurally related intestinal peptide hormones which were purified from a limited number of mammals and are capable of activating the particulate guanylate cyclase-C. Although the biological functions of guanylin and uroguanylin are not yet clarified in detail, they are involved in the r...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1006/bbrc.1999.0719

    authors: Mägert HJ,Hill O,Zucht HD,Martin S,Meyer M,Forssmann WG,Adermann K

    更新日期:1999-05-27 00:00:00

  • Beta-35 is a transferrin-derived inhibitor of angiogenesis and tumor growth.

    abstract::An angiogenesis inhibitor named Beta-35 has been identified and purified from the conditioned medium of mouse pancreatic β cells tumor cells. Beta-35 has a molecular weight of 35 kDa and inhibits DNA synthesis of bovine capillary endothelial cells at a half-maximal concentration of approximately 5 nM. It shows anti-an...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2011.05.045

    authors: Liang G,Butterfield C,Liang J,Birsner A,Folkman J,Shing Y

    更新日期:2011-06-10 00:00:00

  • Preliminary assessment of the C13-side chain 2'-hydroxylase involved in taxol biosynthesis.

    abstract::The biosynthesis of the anticancer drug Taxol in yew (Taxus) species is thought to involve the preliminary formation of the advanced taxane diterpenoid intermediate baccatin III upon which the functionally important N-benzoyl phenylisoserinoyl side chain is subsequently assembled at the C13-O-position. In vivo feeding...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2005.08.119

    authors: Long RM,Croteau R

    更新日期:2005-12-09 00:00:00

  • On the specificity of a bacteriophage-borne endoglycanase for the native capsular polysaccharide produced by Klebsiella pneumoniae SK1 and its derived polymers.

    abstract::The specificity of the endoglycanase associated with the bacteriophage phi SK1 particles was tested on the native capsular polysaccharide produced by Klebsiella pneumoniae serotype SK1 and on three chemically modified polymers derived from it. The primary structure of the SK1 capsular polysaccharide is: [formula: see ...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1006/bbrc.1994.1160

    authors: Cescutti P,Paoletti S

    更新日期:1994-02-15 00:00:00

  • Release of TEA blockade of maxi-K+ channels by isoproterenol.

    abstract::The outward K+ current induced by step depolarization of freshly dispersed myocytes of guinea-pig taenia coli decreased about 80% upon treatment with 3 mM tetraethylammonium chloride (TEA). Isoproterenol (ISO, 2-5 microM) restored it to a large extent. This restoration did not occur in the presence of propranolol (2 m...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1006/bbrc.1994.1664

    authors: Fan SF,Wang S,Kao CY

    更新日期:1994-05-30 00:00:00

  • The association of renin-angiotensin system genes with the progression of hepatocellular carcinoma.

    abstract::Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. Angiogenesis is reported to play a pivotal role in the occurrence, development and metastasis of HCC. The renin-angiotensin system (RAS) is involved in the regulation of angiogenesis. Here, based on the analysis of HCC datasets from Gene Expre...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2015.02.030

    authors: Ye G,Qin Y,Lu X,Xu X,Xu S,Wu C,Wang X,Wang S,Pan D

    更新日期:2015-03-27 00:00:00

  • Lipid fluidity at different regions in LDL and HDL of beta-thalassemia/Hb E patients.

    abstract::Atherosclerosis-related vascular complications in beta-thalassemia/hemoglobin E (beta-thal/Hb E) patients may result from iron induced oxidation of lipoproteins. To identify the specific site of oxidative damage, changes in lipid fluidity at different regions in LDL and HDL particle were investigated using two fluores...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2006.09.106

    authors: Morales NP,Charlermchoung C,Luechapudiporn R,Yamanont P,Fucharoen S,Chantharaksri U

    更新日期:2006-11-24 00:00:00

  • Induction of the lutropin/choriogonadotropin receptor in rat ovary during luteinization.

    abstract::Molecular analysis of the induction of the lutropin/choriogonadotropin receptor during the process of luteinization of the rat ovary was performed. The appearance of receptor binding activity and an immunological analysis of the receptor using Triton solubilized membrane proteins show little receptor present in luteal...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/0006-291x(90)91177-t

    authors: Hansbury MJ,McIlroy PJ,Goldenthal MJ

    更新日期:1990-09-14 00:00:00

  • Do rat hepatic microsomes contain multiple NADPH-supported fatty acid chain elongation pathways or a single pathway?

    abstract::[2-14C]-trans-2-hexadecenoyl CoA (16:1) and [2-14C]-trans-2-cis-8,11,14-eicosatetraenoyl CoA (20:4) were chemically synthesized and employed as competitive substrates for the liver microsomal trans-2-enoyl CoA reductase component of the fatty acid chain elongation system. Both 7.5 microM and 15 microM 20:4 competitive...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/0006-291x(86)91059-4

    authors: Nagi MN,Cook L,Prasad MR,Cinti DL

    更新日期:1986-10-15 00:00:00

  • The modulation of growth by HMBA in PKC overproducing HT29 colon cancer cells.

    abstract::To examine whether protein kinase C (PKC) plays a role in mediating growth inhibitory effects of hexamethylene bisacetamide (HMBA) we compared a control H29 colon cancer cell line to a derivative, HT29-PKC7, that overexpresses high levels of PKC beta 1. We found that although HMBA markedly inhibited the growth of the ...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/0006-291x(91)91262-b

    authors: Choi PM,Weinstein IB

    更新日期:1991-12-16 00:00:00

  • Recombinant human laminin isoforms can support the undifferentiated growth of human embryonic stem cells.

    abstract::Human embryonic stem cells (hESCs) are thought to be a promising cell source for cell transplantation therapy. For such a clinical application, the hESCs should be manipulated using appropriate and qualified materials. In this study, we examined the efficacy of recombinant human laminin (rhLM) isoforms on the undiffer...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2008.07.111

    authors: Miyazaki T,Futaki S,Hasegawa K,Kawasaki M,Sanzen N,Hayashi M,Kawase E,Sekiguchi K,Nakatsuji N,Suemori H

    更新日期:2008-10-10 00:00:00

  • A novel protease inhibitor causes inclusion vacuole reduction and disrupts the intracellular growth of Chlamydia trachomatis.

    abstract::Chlamydia (C.) trachomatis, characterized by a unique biphasic life cycle, is an obligate intracellular bacterial pathogen which is responsible for the highest number of sexually transmitted bacterial infections globally. However, its pathogenic mechanisms have not been fully elucidated because of its unique developme...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2019.05.184

    authors: Zhou Y,Lu X,Huang D,Lu Y,Zhang H,Zhang L,Yu P,Wang F,Wang Y

    更新日期:2019-08-13 00:00:00

  • A label-free impedance-based whole cell assay revealed a new G protein-coupled receptor ligand for mouse microglial cell migration.

    abstract::We report the usefulness of an impedance-based label-free whole cell assay to identify new ligands for G protein-coupled receptors (GPCRs) involved in microglial cell migration. Authentic GPCR ligands were subjected to the impedance-based cell assay in order to examine the responses of ligands for MG5 mouse microglial...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2016.07.119

    authors: Fukano Y,Okino N,Furuya S,Ito M

    更新日期:2016-09-16 00:00:00

  • Transcript decay mediated by RNase III in Borrelia burgdorferi.

    abstract::The causative agent of Lyme disease, Borrelia burgdorferi, requires shifts in gene expression to undergo its natural enzootic cycle between tick and vertebrate hosts. mRNA decay mechanisms play significant roles in governing gene expression in other bacteria, but are not yet characterized in B. burgdorferi. RNase III ...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/j.bbrc.2020.05.201

    authors: Snow S,Bacon E,Bergeron J,Katzman D,Wilhelm A,Lewis O,Syangtan D,Calkins A,Archambault L,Anacker ML,Schlax PJ

    更新日期:2020-08-20 00:00:00

  • Ionophore-induced Cl- transport in human erythrocyte suspensions: a multinuclear magnetic resonance study.

    abstract::To investigate the effect of ionophores on Cl- distribution in human erythrocyte suspensions, we measured the membrane potential by using 19F and 31P NMR methods. Incubation of human erythrocytes with 0.005 mM of the neutral ionophores valinomycin and nonactin resulted in membrane potentials of -21.2 and -17.8 mV in t...

    journal_title:Biochemical and biophysical research communications

    pub_type: 杂志文章

    doi:10.1016/0006-291x(92)90678-e

    authors: Wittenkeller L,Mota de Freitas D,Ramasamy R

    更新日期:1992-04-30 00:00:00