Rapid transfer of overexpressed integral membrane protein from the host membrane into soluble lipid nanodiscs without previous purification.

Abstract:

:Structural and functional characterization of integral membrane proteins in a bilayer environment is strongly hampered by the requirement of detergents for solubilization and subsequent purification, as detergents commonly affect their structure and/or activity. Here, we describe a rapid procedure with minimal exposure to detergent to directly assemble an overexpressed integral membrane protein into soluble lipid nanodiscs prior to purification. This is exemplified with recombinant his-tagged rhodopsin, which is rapidly extracted from its host membrane and directly assembled into membrane scaffold protein (MSP) nanodiscs. We further demonstrate that, even when the MSP was his-tagged as well, partial purification of the rhodopsin-nanodiscs could be achieved exploiting immobilized-metal chromatography. Recoveries of rhodopsin up to 80% were achieved in the purified nanodisc fraction. Over 95% of contaminating membrane protein and his-tagged MSP could be removed from the rhodopsin-nanodiscs using a single Ni2+-affinity chromatography step. This level of purification is amply sufficient for functional studies. We provide evidence that the obtained rhodopsin-nanodisc preparations are fully functional both photochemically and in their ability to bind the cognate G-protein.

journal_name

Biol Chem

journal_title

Biological chemistry

authors

Shirzad-Wasei N,van Oostrum J,Bovee-Geurts PH,Kusters LJ,Bosman GJ,DeGrip WJ

doi

10.1515/hsz-2015-0100

subject

Has Abstract

pub_date

2015-08-01 00:00:00

pages

903-15

issue

8

eissn

1431-6730

issn

1437-4315

pii

/j/bchm.just-accepted/hsz-2015-0100/hsz-2015-0100.

journal_volume

396

pub_type

杂志文章
  • In vitro conversion and seeded fibrillization of posttranslationally modified prion protein.

    abstract::The conversion of the cellular isoform of the prion protein (PrP(C)) into the pathologic isoform (PrP(Sc)) is the key event in prion diseases. To study the conversion process, an in vitro system based on varying the concentration of low amounts of sodium dodecyl sulfate (SDS) has been employed. In the present study, t...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2011.048

    authors: Stöhr J,Elfrink K,Weinmann N,Wille H,Willbold D,Birkmann E,Riesner D

    更新日期:2011-05-01 00:00:00

  • Cooperativity of binding epitopes and receptor chains in the BMP/TGFbeta superfamily.

    abstract::Bone morphogenetic proteins (BMP) are dimeric factors initiating several distinct signaling cascades by binding to two types of transmembrane serine/threonine kinase receptors (BRI and BRII), and are thus regulating several steps in embryonal development and adult tissue homeostasis. BMP-2 contains two symmetrical pai...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2001.149

    authors: Knaus P,Sebald W

    更新日期:2001-08-01 00:00:00

  • Sphingomyelin breakdown in T cells: role in activation, effector functions and immunoregulation.

    abstract::Host T cell activation, a key step in obtaining adaptive immunity against pathogens, is initiated by the binding of the T cell receptor to a foreign antigenic peptide presented by the major histocompatibility complex on the surface of an antigen-presenting cell and, consequently, formation of an immunological synapse....

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/hsz-2014-0282

    authors: Beyersdorf N,Müller N

    更新日期:2015-06-01 00:00:00

  • Role of chitinase-like proteins in cancer.

    abstract::Chitinase-like proteins (CLPs) are lectins combining properties of cytokines and growth factors. Human CLPs include YKL-40, YKL-39 and SI-CLP that are secreted by cancer cells, macrophages, neutrophils, synoviocytes, chondrocytes and other cells. The best investigated CLP in cancer is YKL-40. Serum and plasma levels o...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/hsz-2015-0269

    authors: Kzhyshkowska J,Yin S,Liu T,Riabov V,Mitrofanova I

    更新日期:2016-03-01 00:00:00

  • Molecular function of the prolyl cis/trans isomerase and metallochaperone SlyD.

    abstract::SlyD is a bacterial two-domain protein that functions as a molecular chaperone, a prolyl cis/trans isomerase, and a nickel-binding protein. This review summarizes recent findings about the molecular enzyme mechanism of SlyD. The chaperone function located in one domain of SlyD is involved in twin-arginine translocatio...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/hsz-2013-0137

    authors: Kovermann M,Schmid FX,Balbach J

    更新日期:2013-08-01 00:00:00

  • Functional ribosome biogenesis is a prerequisite for p53 destabilization: impact of chemotherapy on nucleolar functions and RNA metabolism.

    abstract::The production and processing of ribosomal RNA is a complex and well-coordinated nucleolar process for ribosome biogenesis. Progress in understanding nucleolar structure and function has lead to the unexpected discovery of the nucleolus as a highly sensitive sensor of cellular stress and an important regulator of the ...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/hsz-2013-0153

    authors: Burger K,Eick D

    更新日期:2013-09-01 00:00:00

  • The polysaccharide scaffold of PrP 27-30 is a common compound of natural prions and consists of alpha-linked polyglucose.

    abstract::An inert polysaccharide scaffold identified as a 5-15% component of prion rods (PrP 27-30) is unambiguously distinguishable from the N-glycosyl groups and the GPI anchor of PrP, and consists predominantly of 1,4-linked glucose with some branching via 1,4,6-linked glucose. We show that this polysaccharide scaffold is a...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2005.131

    authors: Dumpitak C,Beekes M,Weinmann N,Metzger S,Winklhofer KF,Tatzelt J,Riesner D

    更新日期:2005-11-01 00:00:00

  • Kinetics of autocatalytic zymogen activation measured by a coupled reaction: pepsinogen autoactivation.

    abstract::A kinetic study was performed of a model for an autocatalytic zymogen activation process involving both intra- and intermolecular routes, to which a chromogenic reaction in which the active enzyme acts upon one of its substrates was coupled to continuously monitor the reaction. Kinetic equations describing the evoluti...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2005.080

    authors: Fuentes ME,Varón R,García-Moreno M,Valero E

    更新日期:2005-07-01 00:00:00

  • Functional mapping of the EcoRV DNA methyltransferase by random mutagenesis and screening for catalytically inactive mutants.

    abstract::M.EcoRV is an alpha-adenine DNA methyltransferase. According to structure predictions, the enzyme consists of a catalytic domain, which has a structure similar to all other DNA-methyltransferases, and a smaller DNA-recognition domain. We have investigated this enzyme by random mutagenesis, using error-prone PCR, follo...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/bchm.1998.379.4-5.475

    authors: Friedrich T,Roth M,Helm-Kruse S,Jeltsch A

    更新日期:1998-04-01 00:00:00

  • Interaction of heat shock protein 70 peptide with NK cells involves the NK receptor CD94.

    abstract::Full-length Hsp70 protein (Hsp70) and the C-terminal domain of Hsp70 (Hsp70C) both stimulate the cytolytic activity of naive natural killer (NK) cells against Hsp70-positive tumor target cells. Here, we describe the characterization of Hsp70-NK cell interaction with binding studies using the human NK cell line YT. Bin...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2003.030

    authors: Gross C,Hansch D,Gastpar R,Multhoff G

    更新日期:2003-02-01 00:00:00

  • Bauhinia proteinase inhibitor-based synthetic fluorogenic substrates for enzymes isolated from insect midgut and caterpillar bristles.

    abstract::Bauhinia ungulata factor Xa inhibitor (BuXI) inactivates factor Xa and LOPAP, a prothrombin activator proteinase isolated from the venom of Lonomia obliqua caterpillar bristles. The reactive site of the enzyme-inhibitor interaction was explored to design specific substrates for both enzymes. Methionine is crucial for ...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2003.055

    authors: Andrade SA,Santomauro-Vaz EM,Lopes AR,Chudzinski-Tavassi AM,Juliano MA,Terra WR,Sampaio MU,Sampaio CA,Oliva ML

    更新日期:2003-03-01 00:00:00

  • Nervous system pathology: the fibrin perspective.

    abstract::Studies of extracellular matrix (ECM) biology in the nervous system have mainly focused on laminin, fibronectin and tenascin-R, proteins that are present during nervous system development and normal function. However, during disease, fibrin, which physiologically is not present in the nervous tissue, is detected at ne...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/BC.2002.004

    authors: Akassoglou K,Strickland S

    更新日期:2002-01-01 00:00:00

  • Kinins, receptors, kininases and inhibitors--where did they lead us?

    abstract::Based on studies presented here and other published experiments performed with surviving tissue preparations, with transfected cells and with cells that constitutively express the human angiotensin I converting enzyme ACE and B2 receptors, we concluded the following: ACE inhibitors and other endogenous peptides that r...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/BC.2001.007

    authors: Erdös EG,Marcic BM

    更新日期:2001-01-01 00:00:00

  • Glycine-assisted enhancement of 1,4-beta-d-xylan xylanohydrolase activity at alkaline pH with a pH optimum shift.

    abstract::This is the first report describing the enhancement of xylanase activity by the neutral amino acid glycine. Xylanase activity is increased seven-fold at alkaline pH in the presence of glycine and its pH optimum is shifted from pH 7 to 8 without using any protein engineering techniques. Analysis of the steady-state kin...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2007.007

    authors: Vathipadiekal V,Verma A,Rao M

    更新日期:2007-01-01 00:00:00

  • Homo- and heterodimerization is a common feature of the solute carrier family SLC10 members.

    abstract::The solute carrier family SLC10 consists of seven members, including the bile acid transporters Na+/taurocholate co-transporting polypeptide (NTCP) and apical sodium-dependent bile acid transporter (ASBT), the steroid sulfate transporter SOAT as well as four orphan carriers (SLC10A3, SLC10A4, SLC10A5 and SLC10A7). Pre...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/hsz-2019-0148

    authors: Noppes S,Müller SF,Bennien J,Holtemeyer M,Palatini M,Leidolf R,Alber J,Geyer J

    更新日期:2019-09-25 00:00:00

  • Multifunctional silica nanoparticles for optical and magnetic resonance imaging.

    abstract::The surface of spherical, nonporous silica nanoparticles (SiO2-NPs) was modified with gadolinium (Gd) complexes, fluorophores, and cell-penetrating peptides to achieve multifunctionality on a single particle. The Gd surface concentrations were 9-16 μmol/g resulting in nanomaterials with high local longitudinal and tra...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/hsz-2012-0251

    authors: Joshi R,Feldmann V,Koestner W,Detje C,Gottschalk S,Mayer HA,Sauer MG,Engelmann J

    更新日期:2013-01-01 00:00:00

  • Adjuvant activity of type I interferons.

    abstract::Type I interferons (IFNs) produced primarily by plasmacytoid dendritic cells (pDCs) as part of the innate immune response to infectious agents induce the maturation of myeloid DCs and enhance antigen presentation. Type I IFNs also enhance apoptosis of virus-infected cells, stimulate cross priming and enhanced presenta...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/bc.2008.051

    authors: Tovey MG,Lallemand C,Thyphronitis G

    更新日期:2008-05-01 00:00:00

  • Effects of reducing agents on glutathione metabolism and the function of carotid body chemoreceptor cells.

    abstract::Two current hypotheses of O2 sensing in the carotid body (CB) chemoreceptors suggest participation of oxygen reactive (ROS) species, but they are mechanistically opposed. One postulates that hypoxia decreases ROS levels; the other that hypoxia increases them. Yet, both propose that the ensuing alteration in the cellul...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2004.021

    authors: Gonzalez C,Sanz-Alyayate G,Agapito MT,Obeso A

    更新日期:2004-03-01 00:00:00

  • Three-dimensional cultures modeling premalignant progression of human breast epithelial cells: role of cysteine cathepsins.

    abstract::The expression of the cysteine protease cathepsin B is increased in early stages of human breast cancer.To assess the potential role of cathepsin B in premalignant progression of breast epithelial cells, we employed a 3D reconstituted basement membrane overlay culture model of MCF10A human breast epithelial cells and ...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/hsz-2012-0252

    authors: Mullins SR,Sameni M,Blum G,Bogyo M,Sloane BF,Moin K

    更新日期:2012-12-01 00:00:00

  • The non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase: biochemistry, structure, occurrence and evolution.

    abstract::The non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase catalyses the irreversible reaction of glyceraldehyde-3-phosphate to 3-phosphoglycerate by the reduction of NADP to NADPH. This is in contrast to the extensively analysed phosphorylating glyceraldehyde-3-phosphate dehydrogenases which catalyse the revers...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Habenicht A

    更新日期:1997-12-01 00:00:00

  • Interaction between lipopolysaccharide (LPS), LPS-binding protein (LBP), and planar membranes.

    abstract::The mechanism of interaction of the lipopolysaccharide (LPS)-binding protein, LBP, with differently composed symmetric and asymmetric planar lipid bilayers was investigated in electrical measurements (membrane current, potential, capacitance). From a change of the inner membrane potential difference, binding of LBP to...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2001.052

    authors: Gutsmann T,Haberer N,Carroll SF,Seydel U,Wiese A

    更新日期:2001-03-01 00:00:00

  • Alteration of mosaic methylation of the repeat unit of the human ribosomal RNA genes in lung cancer.

    abstract::We have investigated the methylation status of the repeat unit of the human ribosomal RNA genes in lung cancer. Using a Southern blot analysis approach we have determined that the non-transcribed region of these genes was generally heavily methylated, while the transcribed region was not methylated in either tumor or ...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.1999.010

    authors: Shiraishi M,Sekiguchi A,Chuu YH,Sekiya T

    更新日期:1999-01-01 00:00:00

  • An efficient method for the preparation of long heteroduplex DNA as substrate for mismatch repair by the Escherichia coli MutHLS system.

    abstract::We present a method that allows preparing long DNA containing defined mismatches without the use of gel electrophoretic or chromatographic purification steps. The preparation starts with the synthesis of two PCR products, which are identical except for those positions that will later form the mismatches. One of the PC...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.2002.166

    authors: Thomas E,Pingoud A,Friedhoff P

    更新日期:2002-09-01 00:00:00

  • Contributions of the Ah receptor to bilirubin homeostasis and its antioxidative and atheroprotective functions.

    abstract::Abstract The homeostasis and atheroprotective function of bilirubin could be an appealing model to investigate one of the many physiologic functions of the human aryl hydrocarbon receptor (AhR). Several clinical and epidemiological studies have been carried out on key enzymes generating and eliminating bilirubin (heme...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/BC.2010.065

    authors: Bock KW,Köhle C

    更新日期:2010-06-01 00:00:00

  • Genetically engineered and synthetic allergen derivatives: candidates for vaccination against type I allergy.

    abstract::Type I allergy, a hypersensitivity disease affecting almost 20% of the population worldwide, is based on the IgE recognition of otherwise harmless antigens (i.e., allergens). Allergen-induced crosslink of effector cell-bound IgE antibodies leads to the release of biological mediators and thus to immediate disease symp...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/BC.1999.101

    authors: Valenta R,Vrtala S,Focke-Tejkl M,Bugajska-Schretter,Ball T,Twardosz A,Spitzauer S,Grönlund H,Kraft D

    更新日期:1999-07-01 00:00:00

  • Tapasin and other chaperones: models of the MHC class I loading complex.

    abstract::MHC (major histocompatibility complex) class I molecules bind intracellular virus-derived peptides in the endoplasmic reticulum (ER) and present them at the cell surface to cytotoxic T lymphocytes. Peptide-free class I molecules at the cell surface, however, could lead to aberrant T cell killing. Therefore, cells ensu...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/BC.2004.100

    authors: Wright CA,Kozik P,Zacharias M,Springer S

    更新日期:2004-09-01 00:00:00

  • The YidC/Oxa1/Alb3 protein family: common principles and distinct features.

    abstract::The members of the YidC/Oxa1/Alb3 protein family are evolutionary conserved in all three domains of life. They facilitate the insertion of membrane proteins into bacterial, mitochondrial, and thylakoid membranes and have been implicated in membrane protein folding and complex formation. The major classes of substrates...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/hsz-2012-0199

    authors: Saller MJ,Wu ZC,de Keyzer J,Driessen AJ

    更新日期:2012-11-01 00:00:00

  • Functional characterization of atherosclerosis-associated Ser128Arg and Leu554Phe E-selectin mutations.

    abstract::The cellular adhesion molecule E-selectin is expressed on activated endothelial cells, and is involved in the process of adherence of blood cells to vessel endothelium in inflammatory events such as atherosclerosis. In a recent study we found a Ser128Arg mutation in the EGF domain as well as a Leu554Phe mutation in th...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/BC.1999.082

    authors: Wenzel K,Stahn R,Speer A,Denner K,Gläser C,Affeldt M,Moobed M,Scheer A,Baumann G,Felix SB

    更新日期:1999-06-01 00:00:00

  • Annexins - insights from knockout mice.

    abstract::Annexins are a highly conserved protein family that bind to phospholipids in a calcium (Ca2+) - dependent manner. Studies with purified annexins, as well as overexpression and knockdown approaches identified multiple functions predominantly linked to their dynamic and reversible membrane binding behavior. However, mos...

    journal_title:Biological chemistry

    pub_type: 杂志文章,评审

    doi:10.1515/hsz-2016-0168

    authors: Grewal T,Wason SJ,Enrich C,Rentero C

    更新日期:2016-10-01 00:00:00

  • Cis-regulatory sequences from the first intron of the rat glutamine synthetase gene are involved in hepatocyte specific expression of the enzyme.

    abstract::In order to identify regulatory elements involved in the hepatocyte specific expression of the enzyme glutamine synthetase [GS (E.C. 6.3.1.2)] we analyzed the first intron of the rat GS gene. A sequence analysis detected clusters of potential transcription factor binding sites in regions that are hypersensitive for DN...

    journal_title:Biological chemistry

    pub_type: 杂志文章

    doi:10.1515/bchm.1997.378.1.11

    authors: Gaunitz F,Gaunitz C,Papke M,Gebhardt R

    更新日期:1997-01-01 00:00:00