Cross-linking of dimeric CitS and GltS transport proteins.

Abstract:

:CitS of Klebsiella pneumoniae and GltS of Escherichia coli are Na+-dependent secondary transporters from different families that are believed to share the same fold and quaternary structure. A 10 kDa protein tag (Biotin Acceptor Domain [BAD]) was fused to the N-terminus of both proteins (CitS-BAD1 and GltS-BAD1, respectively) and inserted in the central cytoplasmic loop that connects the two halves of the proteins (CitS-BAD260 and GltS-BAD206). Both CitS constructs and GltS-BAD206 were produced and shown to be active transporters, but GltS-BAD1 could not be detected in the membrane. Distance relationships in the complexes were studied by cross-linking studies. Both CitS constructs were shown to be in the dimeric state after purification in detergent by cross-linking with glutaraldehyde. The concentration of glutaraldehyde resulting in 50% cross-linking was significantly higher for CitS-BAD1 than for CitS and CitS-BAD260. Remarkably, GltS and GltS-BAD260 were not cross-linked by glutaraldehyde because of the lack of productive reactive sites. Cross-linking of GltS was observed when the N-terminal 46 residues of CitS with or without BAD at the N-terminus were added to the N-terminus of GltS. The stretch of 46 residues contains the first transmembrane segment of CitS that is missing in the GltS structure. The data support an orientation of the monomers in the dimer with the N-termini close to the dimer interface and the central cytoplasmic loops far away at the ends of the long axis of the dimer structure in a view perpendicular to the membrane.

journal_name

Mol Membr Biol

authors

Krupnik T,Dobrowolski A,Lolkema JS

doi

10.3109/09687688.2011.581252

subject

Has Abstract

pub_date

2011-08-01 00:00:00

pages

243-53

issue

5

eissn

0968-7688

issn

1464-5203

journal_volume

28

pub_type

杂志文章
  • In vivo molecular imaging using nanomaterials: general in vivo characteristics of nano-sized reagents and applications for cancer diagnosis.

    abstract::Nanoparticles present a new collection of contrast agents for the field of in vivo molecular imaging. This review focuses on promising molecular imaging probes for optical and magnetic resonance imaging based on four representative nanomaterial(s) platforms: quantum dots, upconversion phosphors, superparamagnetic iron...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.3109/09687688.2010.481640

    authors: Rosenblum LT,Kosaka N,Mitsunaga M,Choyke PL,Kobayashi H

    更新日期:2010-10-01 00:00:00

  • Ion channels of glutamate receptors: structural modeling.

    abstract::Ionotropic glutamate receptors belong to the superfamily of P-loop channels as well as K(+), Na(+), and Ca(2+) channels. However, the structural similarity between ion channels of the glutamate receptors and K(+) channels is a matter of discussion. The aim of this study was to analyze differences between the structure...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/09687860601008806

    authors: Tikhonov DB

    更新日期:2007-03-01 00:00:00

  • Patterns and processes in the evolution of the eukaryotic endomembrane system.

    abstract::The eukaryotic endomembrane system (ES) is served by hundreds of dedicated proteins. Experimental characterization of the ES-associated molecular machinery in several model eukaryotes complemented by a recent progress in phylogenomics and comparative genomics have revealed a conserved complex core of the machinery tha...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.3109/09687688.2010.521201

    authors: Elias M

    更新日期:2010-11-01 00:00:00

  • Slippery signaling: Palmitoylation-dependent control of neuronal kinase localization and activity.

    abstract::Modification of proteins with the lipid palmitate, a process called palmitoylation, is important for the normal function of neuronal cells. However, most attention has focused on how palmitoylation regulates the targeting and trafficking of neurotransmitter receptors and non-enzymatic scaffold proteins. In this review...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.1080/09687688.2016.1182652

    authors: Montersino A,Thomas GM

    更新日期:2015-08-01 00:00:00

  • Sphingomyelin chain length influences the distribution of GPI-anchored proteins in rafts in supported lipid bilayers.

    abstract::Glycosyl-phosphatidylinositol (GPI)-anchored proteins are enriched in cholesterol- and sphingolipid-rich lipid rafts within the membrane. Rafts are known to have roles in cellular organization and function, but little is understood about the factors controlling the distribution of proteins in rafts. We have used atomi...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/09687860601127770

    authors: Garner AE,Smith DA,Hooper NM

    更新日期:2007-05-01 00:00:00

  • Purification and properties of the Escherichia coli nucleoside transporter NupG, a paradigm for a major facilitator transporter sub-family.

    abstract::NupG from Escherichia coli is the archetype of a family of nucleoside transporters found in several eubacterial groups and has distant homologues in eukaryotes, including man. To facilitate investigation of its molecular mechanism, we developed methods for expressing an oligohistidine-tagged form of NupG both at high ...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/09687860400003941

    authors: Xie H,Patching SG,Gallagher MP,Litherland GJ,Brough AR,Venter H,Yao SY,Ng AM,Young JD,Herbert RB,Henderson PJ,Baldwin SA

    更新日期:2004-09-01 00:00:00

  • Role of glutamate-269 in the lactose permease of Escherichia coli.

    abstract::Glu-269, which is located on the hydrophilic face of putative helix VIII in the lactose permease of Escherichia coli, has been replaced with Asp, Gln or Cys by oligonucleotide-directed, site specific mutagenesis. Cells expressing Asp-269 permease exhibit no lactose accumulation or lactose-induced H+ translocation, but...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687689409161024

    authors: Ujwal ML,Sahin-Tóth M,Persson B,Kaback HR

    更新日期:1994-01-01 00:00:00

  • The effect of single cerebroside compounds on activation of BKCa channels.

    abstract::We have previously shown that a mixture of cerebrosides obtained from dried tubers of herb Typhonium giganteum Engl. plays a neuroprotective role in the ischemic brain through its effect on activation of BK(Ca) channels. It is very curious to know whether a single pure cerebroside compound could activate the BK(Ca) ch...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687688.2010.538731

    authors: Xu H,Qi J,Wang G,Deng H,Qi Z

    更新日期:2011-02-01 00:00:00

  • Percolation properties of two-component, two-phase phospholipid bilayers.

    abstract::Temperature-composition phase diagrams of binary lipid mixtures under conditions of constant pressure and excess water show that component immiscibility is the rule rather than the exception in these systems. Phase immiscibility is particularly noted in the solid phase but several cases are known where immiscibility i...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687689509038493

    authors: Vaz WL

    更新日期:1995-01-01 00:00:00

  • In vitro folding of a membrane protein: effect of denaturation and renaturation on substrate binding by the lactose permease of Escherichia coli.

    abstract::Site-directed mutagenesis and site-directed fluorescence spectroscopy demonstrate that Cys148 interacts hydrophobically with the galactosyl moiety of substrates of the lactose permease of Escherichia coli. By taking advantage of the finding that labelling of single-Cys148 permease with the thiol-specific fluorophore 2...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687689809027513

    authors: He MM,Kaback HR

    更新日期:1998-01-01 00:00:00

  • The Equilibrative Nucleoside Transporter (ENT1) can be phosphorylated at multiple sites by PKC and PKA.

    abstract::Equilibrative Nucleoside Transporters (SLC29) are a family of proteins that transport nucleosides, nucleobases and nucleoside analogue drugs across cellular membranes. ENT1 is expressed ubiquitously in mammalian tissues and responsible for a significant portion of nucleoside analog drug uptake in humans. Despite the i...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687688.2011.604861

    authors: Reyes G,Nivillac NM,Karim MZ,Desouza L,Siu KW,Coe IR

    更新日期:2011-09-01 00:00:00

  • The Amt/Mep/Rh family of ammonium transport proteins.

    abstract::The Amt/Mep/Rh family of integral membrane proteins comprises ammonium transporters of bacteria, archaea and eukarya, as well as the Rhesus proteins found in animals. They play a central role in the uptake of reduced nitrogen for biosynthetic purposes, in energy metabolism, or in renal excretion. Recent structural inf...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.1080/09687680701388423

    authors: Andrade SL,Einsle O

    更新日期:2007-09-01 00:00:00

  • Multiple levels of Notch signal regulation (review).

    abstract::Notch is a vitally important signalling receptor controlling cell fate determination and pattern formation in numerous ways during development of both invertebrate and vertebrate species. An intriguing pathway for the Notch signal has emerged where, after ligand-dependent proteolysis, an intracellular fragment of the ...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.1080/09687680110112929

    authors: Baron M,Aslam H,Flasza M,Fostier M,Higgs JE,Mazaleyrat SL,Wilkin MB

    更新日期:2002-01-01 00:00:00

  • Regulation of the calpain-calpastatin system by membranes (review).

    abstract::Calpain, an intracellular calcium-dependent protease, is activated at cell membranes and cleaves cytoskeletal and submembranous proteins. Calpain is inferred to be a calcium-dependent regulator for cytoskeletal reorganization. Calpastatin, an endogenous calpain inhibitor, inhibits not only the proteolytic activity of ...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.3109/09687689609160599

    authors: Kawasaki H,Kawashima S

    更新日期:1996-10-01 00:00:00

  • Functional engineered channels and pores (Review).

    abstract::Significant progress has been made in membrane protein engineering over the last 5 years, based largely on the re-design of existing scaffolds. Engineering techniques that have been employed include direct genetic engineering, both covalent and non-covalent modification, unnatural amino acid mutagenesis and total synt...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.1080/09687680410001716853

    authors: Bayley H,Jayasinghe L

    更新日期:2004-07-01 00:00:00

  • Topological and segmental phylogenetic analyses of the anion exchanger (band 3) family of transporters.

    abstract::Eleven sequenced anion exchanger (AE; band 3) proteins, including five AE1, four AE2 and two AE3 proteins, comprise the anion exchanger family (AEF) of homologous proteins. Eliminating the rat and rabbit proteins that are nearly identical to the corresponding mouse proteins, seven dissimilar members of this family wer...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687689509027507

    authors: Espanol MJ,Saier MH Jr

    更新日期:1995-04-01 00:00:00

  • Probing the contacts of a low-affinity substrate with a membrane-embedded transport protein using 1H-13C cross-polarisation magic-angle spinning solid-state NMR.

    abstract::Solid-state NMR combined with sample deuteration was used to probe the proximity of the low-affinity substrate D-glucose to its binding site within the Escherichia coli sugar transport protein GalP. Samples of E. coli inner membranes with amplified expression of GalP were incubated in D(2)O with D-[(13)C(6)]glucose an...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687688.2012.743193

    authors: Patching SG,Henderson PJ,Sharples DJ,Middleton DA

    更新日期:2013-03-01 00:00:00

  • The substrate specificity of the human ADP/ATP carrier AAC1.

    abstract::The mitochondrial ADP/ATP carrier imports ADP from the cytosol into the mitochondrial matrix for its conversion to ATP by ATP synthase and exports ATP out of the mitochondrion to replenish the eukaryotic cell with chemical energy. Here the substrate specificity of the human mitochondrial ADP/ATP carrier AAC1 was deter...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687688.2012.745175

    authors: Mifsud J,Ravaud S,Krammer EM,Chipot C,Kunji ER,Pebay-Peyroula E,Dehez F

    更新日期:2013-03-01 00:00:00

  • Membrane topology of the cyanobacterial bicarbonate transporter, SbtA, and identification of potential regulatory loops.

    abstract::The transporter SbtA is a high affinity Na+-dependent HCO3- uptake system present in a majority of cyanobacterial clades. It functions in conjunction with CO2 uptake systems and other HCO3- uptake systems to allow cyanobacteria to accumulate high levels of HCO3- used to support efficient photosynthetic CO2 fixation vi...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687688.2011.593049

    authors: Price GD,Shelden MC,Howitt SM

    更新日期:2011-08-01 00:00:00

  • Dynamic and cellular interactions of nanoparticles in vascular-targeted drug delivery (review).

    abstract::Vascular-targeted drug delivery systems could provide more efficient and effective pharmaceutical interventions for treating a variety of diseases including cardiovascular, pulmonary, inflammatory, and malignant disorders. However, several factors must be taken into account when designing these systems. The diverse bl...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.3109/09687688.2010.499548

    authors: Huang RB,Mocherla S,Heslinga MJ,Charoenphol P,Eniola-Adefeso O

    更新日期:2010-08-01 00:00:00

  • Peptidase allergen Der p 1 initiates apoptosis of epithelial cells independently of tight junction proteolysis.

    abstract::Loss of epithelial cell polarity, which can arise following disruption of tight junctions (TJs), is a precursor to the care-fully orchestrated removal of moribund cells from epithelia in apoptosis. Ordinarily, this cycle of events has minimally disruptive effects on the function of the epithelial barrier, but some age...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/0968768021000061150

    authors: Baker SF,Yin Y,Runswick SK,Stewart GA,Thompson PJ,Garrod DR,Robinson C

    更新日期:2003-01-01 00:00:00

  • Intracellular pH regulation in ventral horn neurones cultured from embryonic rat spinal cord.

    abstract::Intracellular pH was measured with the pH-sensitive fluorescent probe BCECF in spinal cord neurones cultured from rat embryos. At an external pH of 7.3, the average steady-state pHi was 7.18 +/- 0.03 (SEM, n = 97) and 7.02 +/- 0.01 (n = 221) in HEPES-buffered and in bicarbonate-buffered medium, respectively. In both e...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/09687680050117129

    authors: Brechenmacher C,Rodeau JL

    更新日期:2000-04-01 00:00:00

  • Structural and dynamic studies of the gamma-M4 trans-membrane domain of the nicotinic acetylcholine receptor.

    abstract::A structural characterization of a synthetic peptide corresponding to the fourth transmembrane domain (M4-TMD) of the gamma-subunit of the nicotinic acetylcholine receptor from Torpedo californica has been undertaken. Solid-state NMR and CD spectroscopy studies indicate that upon reconstitution into lipid vesicles or ...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/09687860500370653

    authors: Williamson PT,Zandomeneghi G,Barrantes FJ,Watts A,Meier BH

    更新日期:2005-11-01 00:00:00

  • Interactions of the monomeric and dimeric flavones apigenin and amentoflavone with the plasma membrane of L929 cells; a fluorescence study.

    abstract::Flavonoids are ubiquitous polyphenolic compounds, found in vascular plants, which are endowed with a large variety of biological effects. Some of these effects have been assumed to result from interactions with the cell plasma membrane. In order to investigate the nature of these interactions a fluorescence study was ...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/096876899294616

    authors: Lenne-Gouverneur AF,Lobstein A,Haan-Archipoff G,Duportail G,Anton R,Kuhry JG

    更新日期:1999-04-01 00:00:00

  • Structural and functional divergence of the newly identified GtrIc from its Gtr family of conserved Shigella flexneri serotype-converting glucosyltransferases.

    abstract::Glucosyltransferases (Gtrs) and O-acetyltransferase (Oac) are integral membrane proteins embedded within the cytoplasmic membrane of Shigella flexneri. Gtrs and Oac are responsible for unidirectional host serotype conversion by altering the epitopic properties of the bacterial surface lipopolysaccharide (LPS) O-antige...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687680903552250

    authors: Ramiscal RR,Tang SS,Korres H,Verma NK

    更新日期:2010-04-01 00:00:00

  • Location and membrane sources for autophagosome formation - from ER-mitochondria contact sites to Golgi-endosome-derived carriers.

    abstract::Recent advances have revealed much about the signaling events and molecular components associated with autophagy. Although it is clear that there are multiple points of intersection and connection between autophagy and known vesicular membrane transport processes between membrane compartments, autophagy is modulated b...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687688.2013.850178

    authors: Chan SN,Tang BL

    更新日期:2013-12-01 00:00:00

  • Solubilization of high affinity G-protein-coupled serotonin1A receptors from bovine hippocampus using pre-micellar CHAPS at low concentration.

    abstract::The serotonin1A (5-HT1A) receptors are members of a superfamily of seven transmembrane domain receptors that couple to G-proteins. They appear to be involved in various behavioural and cognitive functions. This paper reports an efficient strategy to solubilize 5-HT1A receptors from bovine hippocampal membranes using t...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.1080/09687680210149586

    authors: Chattopadhyay A,Harikumar KG,Kalipatnapu S

    更新日期:2002-07-01 00:00:00

  • TGN38 cycles via the basolateral membrane of polarized Caco-2 cells.

    abstract::TGN38 is a heavily glycosylated, type I integral membrane protein which is predominantly localized to the trans Golgi network (TGN), but which constitutively traffics between the TGN and the cell surface. The trafficking of TGN38 has been extensively studied in non-polarized cells, and a short, tyrosine-based, peptide...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687689809074524

    authors: Reaves BJ,Roquemore EP,Luzio JP,Banting G

    更新日期:1998-07-01 00:00:00

  • A virtual high-throughput screening approach to the discovery of novel inhibitors of the bacterial leucine transporter, LeuT.

    abstract::Membrane proteins are intrinsically involved in both human and pathogen physiology, and are the target of 60% of all marketed drugs. During the past decade, advances in the studies of membrane proteins using X-ray crystallography, electron microscopy and NMR-based techniques led to the elucidation of over 250 unique m...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章

    doi:10.3109/09687688.2012.710341

    authors: Simmons KJ,Gotfryd K,Billesbølle CB,Loland CJ,Gether U,Fishwick CW,Johnson AP

    更新日期:2013-03-01 00:00:00

  • Lipid rafts in T cell receptor signalling .

    abstract::The molecular events and the protein components that are involved in signalling by the T cell receptor (TCR) for antigen have been extensively studied. Activation of signalling cascades following TCR stimulation depends on the phosphorylation of the receptor by the tyrosine kinase Lck, which localizes to the cytoplasm...

    journal_title:Molecular membrane biology

    pub_type: 杂志文章,评审

    doi:10.1080/09687860500453673

    authors: Kabouridis PS

    更新日期:2006-01-01 00:00:00