Forming giant vesicles with controlled membrane composition, asymmetry, and contents.

Abstract:

:Growing knowledge of the key molecular components involved in biological processes such as endocytosis, exocytosis, and motility has enabled direct testing of proposed mechanistic models by reconstitution. However, current techniques for building increasingly complex cellular structures and functions from purified components are limited in their ability to create conditions that emulate the physical and biochemical constraints of real cells. Here we present an integrated method for forming giant unilamellar vesicles with simultaneous control over (i) lipid composition and asymmetry, (ii) oriented membrane protein incorporation, and (iii) internal contents. As an application of this method, we constructed a synthetic system in which membrane proteins were delivered to the outside of giant vesicles, mimicking aspects of exocytosis. Using confocal fluorescence microscopy, we visualized small encapsulated vesicles docking and mixing membrane components with the giant vesicle membrane, resulting in exposure of previously encapsulated membrane proteins to the external environment. This method for creating giant vesicles can be used to test models of biological processes that depend on confined volume and complex membrane composition, and it may be useful in constructing functional systems for therapeutic and biomaterials applications.

authors

Richmond DL,Schmid EM,Martens S,Stachowiak JC,Liska N,Fletcher DA

doi

10.1073/pnas.1016410108

subject

Has Abstract

pub_date

2011-06-07 00:00:00

pages

9431-6

issue

23

eissn

0027-8424

issn

1091-6490

pii

1016410108

journal_volume

108

pub_type

杂志文章
  • The beta subunit of the high-conductance calcium-activated potassium channel contributes to the high-affinity receptor for charybdotoxin.

    abstract::Transient expression of either alpha or alpha + beta subunits of the high-conductance Ca2+-activated K+ (maxi-K) channel has been achieved in COS-1 cells. Expression has been studied using charybdotoxin (ChTX), a peptidyl inhibitor that binds in the pore on the alpha subunit. Although some properties of monoiodotyrosi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.94.7.2853

    authors: Hanner M,Schmalhofer WA,Munujos P,Knaus HG,Kaczorowski GJ,Garcia ML

    更新日期:1997-04-01 00:00:00

  • Protein kinase A-anchoring (AKAP) domains in brefeldin A-inhibited guanine nucleotide-exchange protein 2 (BIG2).

    abstract::Like other guanine nucleotide-exchange proteins (GEPs) that activate ADP-ribosylation factor (ARF) GTPases, brefeldin A-inhibited GEP2, BIG2, contains an approximately 200-aa Sec7 domain that is responsible for this catalytic activity and its inhibition by brefeldin A. The Sec7 domain is located near the center of the...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0337678100

    authors: Li H,Adamik R,Pacheco-Rodriguez G,Moss J,Vaughan M

    更新日期:2003-02-18 00:00:00

  • Identification of T6SS-dependent effector and immunity proteins by Tn-seq in Vibrio cholerae.

    abstract::Type VI protein secretion system (T6SS) is important for bacterial competition through contact-dependent killing of competitors. T6SS delivers effectors to neighboring cells and corresponding antagonistic proteins confer immunity against effectors that are delivered by sister cells. Although T6SS has been found in mor...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1222783110

    authors: Dong TG,Ho BT,Yoder-Himes DR,Mekalanos JJ

    更新日期:2013-02-12 00:00:00

  • Desmosomal adhesion inhibits invasive behavior.

    abstract::Recent studies of human disease and transgenic animal experiments have clearly demonstrated the importance of desmosomes in normal tissue architecture. Furthermore, desmosomal components are down-regulated in certain types of carcinomas, suggesting a possible role for desmosomes in suppression of invasion and metastas...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.95.14.8064

    authors: Tselepis C,Chidgey M,North A,Garrod D

    更新日期:1998-07-07 00:00:00

  • Loss of epidermal growth factor requirement and malignant transformation.

    abstract::Serum provides growth factors that regulate and limit the growth of normal cells in tissue culture. Animal cells that are malignantly transformed usually exhibit diminished serum requirements for growth in culture. We have used a defined, serum-free medium to determine which of these growth factors becomes dispensable...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.76.8.3937

    authors: Cherington PV,Smith BL,Pardee AB

    更新日期:1979-08-01 00:00:00

  • Reaction sequence and molecular mass of a Cl(-)-translocating P-type ATPase.

    abstract::The basolateral membranes of Aplysia californica foregut absorptive cells contain both Cl(-)-stimulated ATPase and ATP-dependent Cl- transport activities, and each was inhibited by orthovanadate. Both of these orthovanadate-sensitive activities were reconstituted into proteoliposomes. The reaction sequence kinetics we...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.17.7970

    authors: Gerencser GA,Zelezna B

    更新日期:1993-09-01 00:00:00

  • A geometric identity for Pappus' theorem.

    abstract::An expression in the exterior algebra of a Peano space yielding Pappus' theorem was originally given by Doubilet, Rota, and Stein [Doubilet, P., Rota, G.-C. & Stein, J. (1974) Stud. Appl. Math. 8, 185-216]. Motivated by an identity of Rota, I give an identity in a Grassmann-Cayley algebra of step 3, involving joins an...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.91.8.2909

    authors: Hawrylycz M

    更新日期:1994-04-12 00:00:00

  • Expression profiles of multiple genes in single neurons of Alzheimer's disease.

    abstract::Many changes have been described in the brains of Alzheimer's disease (AD) patients, including loss of neurons and formation of senile plaques and neurofibrillary tangles. The molecular mechanisms underlying these pathologies are unclear. Northern blot, dot-blot, and reverse transcription-coupled PCR analyses have dem...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.95.16.9620

    authors: Chow N,Cox C,Callahan LM,Weimer JM,Guo L,Coleman PD

    更新日期:1998-08-04 00:00:00

  • Social, institutional, and knowledge mechanisms mediate diverse ecosystem service benefits from coral reefs.

    abstract::Ecosystem services are supplied by nature but, by definition, are received by people. Ecosystem service assessments, intended to influence the decisions people make regarding their interactions with nature, need to understand how people benefit from different ecosystem services. A critical question is therefore, What ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1413473111

    authors: Hicks CC,Cinner JE

    更新日期:2014-12-16 00:00:00

  • Heat-shock protein 104 expression is sufficient for thermotolerance in yeast.

    abstract::In all organisms, mild heat pretreatments induce tolerance to high temperatures. In the yeast Saccharomyces cerevisiae, such pretreatments strongly induce heat-shock protein (Hsp) 104, and hsp104 mutations greatly reduce high-temperature survival, indicating Hsp1O4 plays a critical role in induced thermotolerance. Sur...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.11.5301

    authors: Lindquist S,Kim G

    更新日期:1996-05-28 00:00:00

  • Autologous engineering of cartilage.

    abstract::Treatment of full-thickness damage to hyaline cartilage is hampered by the limited availability of autologous healthy cartilage and the lengthy, cost-prohibitive cell isolation and expansion steps associated with autologous cartilage implantation (ACI). Here we report a strategy for de novo engineering of ectopic auto...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0907774107

    authors: Emans PJ,van Rhijn LW,Welting TJ,Cremers A,Wijnands N,Spaapen F,Voncken JW,Shastri VP

    更新日期:2010-02-23 00:00:00

  • Collagen synthesis: a disulfide-linked collagen precursor in chick bone.

    abstract::Chick calvaria labeled in vitro contain a triple-stranded collagen precursor that contains two pro alpha1 and one pro alpha2 chains. All chains contain cystine and are linked by S-S bonds. Ultracentrifugal analysis was used to measure the relative size of the disulfide-linked units. It is proposed that the S-S links h...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.70.10.2993

    authors: Fessler LI,Burgeson RE,Morris NP,Fessler JH

    更新日期:1973-10-01 00:00:00

  • Soft X-ray tomography of phenotypic switching and the cellular response to antifungal peptoids in Candida albicans.

    abstract::The opportunistic pathogen Candida albicans can undergo phenotypic switching between a benign, unicellular phenotype and an invasive, multicellular form that causes candidiasis. Increasingly, strains of Candida are becoming resistant to antifungal drugs, making the treatment of candidiasis difficult, especially in imm...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0906145106

    authors: Uchida M,McDermott G,Wetzler M,Le Gros MA,Myllys M,Knoechel C,Barron AE,Larabell CA

    更新日期:2009-11-17 00:00:00

  • A 170-kDa membrane-bound protease is associated with the expression of invasiveness by human malignant melanoma cells.

    abstract::Malignant spreading of cancer cells requires cell surface proteases that cleave the crosslinked collagenous matrix of connective tissues. From correlating the morphologically defined invasiveness of tumor cells with the presence of specific membrane-associated proteases, we have identified a malignant human melanoma c...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.87.21.8296

    authors: Aoyama A,Chen WT

    更新日期:1990-11-01 00:00:00

  • A thermodynamic model of regulation: modulation of redox equilibria in camphor monoxygenase.

    abstract::Regulation of biological phenomena occurs in all types of systems, being manifested in many different reaction types, from allosteric behavior in proteins, through modulation in energy and information transfer, to the control of growth and differentiation in cells, organelles, and organisms. In this communication, a m...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.73.4.1078

    authors: Sligar SG,Gunsalus IC

    更新日期:1976-04-01 00:00:00

  • Wild-type p53 controls cell motility and invasion by dual regulation of MET expression.

    abstract::Recent observations suggest that p53 mutations are responsible not only for growth of primary tumors but also for their dissemination. However, mechanisms involved in p53-mediated control of cell motility and invasion remain poorly understood. By using the primary ovarian surface epithelium cell culture, we show that ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1017536108

    authors: Hwang CI,Matoso A,Corney DC,Flesken-Nikitin A,Körner S,Wang W,Boccaccio C,Thorgeirsson SS,Comoglio PM,Hermeking H,Nikitin AY

    更新日期:2011-08-23 00:00:00

  • Specificity in the activation and control of transcription factor E2F-dependent apoptosis.

    abstract::Previous work has demonstrated a role for the E2F1 gene product in signaling apoptosis, both as a result of the deregulation of the Rb/E2F pathway as well as in response to DNA damage. We now show that the ability of cells to suppress the apoptotic potential of E2F1, as might occur during the course of normal cellular...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1831408100

    authors: Hallstrom TC,Nevins JR

    更新日期:2003-09-16 00:00:00

  • Ultrastructural localization of tyrosine hydroxylase in noradrenergic neurons of brain.

    abstract::Tyrosine hydroxylase (EC 1.14.16.2), the enzyme catalyzing the rate-limiting step in catecholamine biosynthesis, was localized by electron microscopy within noradrenergic neurons of the nucleus locus coeruleus of the rat brain with a specific antibody to tyrosine hydroxylase labeled by the peroxidase-antiperoxidase im...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.72.2.659

    authors: Pickel VM,Joh TH,Reis DJ

    更新日期:1975-02-01 00:00:00

  • Reduced amounts of cartilage collagen fibrils and growth plate anomalies in transgenic mice harboring a glycine-to-cysteine mutation in the mouse type II procollagen alpha 1-chain gene.

    abstract::We have generated transgenic mice harboring a glycine-to-cysteine mutation in residue 85 of the triple helical domain of mouse type II collagen. The offspring of different founders displayed a phenotype of severe chondrodysplasia characterized by short limbs and trunk, cranio-facial deformities, and cleft palate. The ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.88.21.9648

    authors: Garofalo S,Vuorio E,Metsaranta M,Rosati R,Toman D,Vaughan J,Lozano G,Mayne R,Ellard J,Horton W

    更新日期:1991-11-01 00:00:00

  • Social isolation uncovers a circuit underlying context-dependent territory-covering micturition.

    abstract::The release of urine, or micturition, serves a fundamental physiological function and, in many species, is critical for social communication. In mice, the pattern of urine release is modulated by external and internal factors and transmitted to the spinal cord via the pontine micturition center (PMC). Here, we exploit...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.2018078118

    authors: Hyun M,Taranda J,Radeljic G,Miner L,Wang W,Ochandarena N,Huang KW,Osten P,Sabatini BL

    更新日期:2021-01-05 00:00:00

  • Identification of murine homologues of the Drosophila son of sevenless gene: potential activators of ras.

    abstract::Several findings suggest that signals from tyrosine kinases are transduced, at least in part, through ras proteins. These findings include (i) blockage of the transforming activity of constitutively active tyrosine kinases by inhibiting ras function and (ii) genetic screens in Caenorhabditis elegans and in Drosophila ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.89.14.6511

    authors: Bowtell D,Fu P,Simon M,Senior P

    更新日期:1992-07-15 00:00:00

  • Maturation of thylakoid lumen proteins proceeds post-translationally through an intermediate in vivo.

    abstract::Many thylakoid lumen proteins are synthesized outside the chloroplast as larger molecular weight precursors and then processed to their mature size during transport to the lumenal space. We have examined the post-translational processing of thylakoid lumen proteins in vivo by pulse-radiolabeling experiments with Chlam...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.5.1862

    authors: Howe G,Merchant S

    更新日期:1993-03-01 00:00:00

  • A functional interleukin 12 receptor complex is composed of two beta-type cytokine receptor subunits.

    abstract::We have identified a cDNA from a human phytohemagglutinin-activated lymphoblast library encoding a protein that binds 125I-labeled human interleukin 12 (125I-huIL-12) with a Kd of about 5 nM when expressed in COS-7 cells. When coexpressed in COS-7 cells with the previously identified IL-12 beta receptor (IL-12R beta) ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.24.14002

    authors: Presky DH,Yang H,Minetti LJ,Chua AO,Nabavi N,Wu CY,Gately MK,Gubler U

    更新日期:1996-11-26 00:00:00

  • Laurasian migration explains Gondwanan disjunctions: evidence from Malpighiaceae.

    abstract::Explanations for biogeographic disjunctions involving South America and Africa typically invoke vicariance of western Gondwanan biotas or long distance dispersal. These hypotheses are problematical because many groups originated and diversified well after the last known connection between Africa and South America (app...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.102175899

    authors: Davis CC,Bell CD,Mathews S,Donoghue MJ

    更新日期:2002-05-14 00:00:00

  • Simple and complex nuclear loci created by newly transferred chloroplast DNA in tobacco.

    abstract::Transfer of organelle DNA into the nuclear genome has been significant in eukaryotic evolution, because it appears to be the origin of many nuclear genes. Most studies on organelle DNA transfer have been restricted to evolutionary events but experimental systems recently became available to monitor the process in real...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0400853101

    authors: Huang CY,Ayliffe MA,Timmis JN

    更新日期:2004-06-29 00:00:00

  • Salt bridge integrates GPCR activation with protein trafficking.

    abstract::G protein-coupled receptors (GPCRs) play central roles in almost all physiological functions; mutations in GPCRs are responsible for more than 30 disorders. There is a great deal of information about GPCR structure but little information that directly relates structure to protein trafficking or to activation. The gona...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0914261107

    authors: Janovick JA,Conn PM

    更新日期:2010-03-02 00:00:00

  • Ex vivo pretreatment of bone marrow mononuclear cells with endothelial NO synthase enhancer AVE9488 enhances their functional activity for cell therapy.

    abstract::Bone marrow mononuclear cells (BMC) from patients with ischemic cardiomyopathy (ICMP) show a reduced neovascularization capacity in vivo. NO plays an important role in neovascularization, and NO bioavailability is typically reduced in patients with ICMP. We investigated whether the impaired neovascularization capacity...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0604144103

    authors: Sasaki K,Heeschen C,Aicher A,Ziebart T,Honold J,Urbich C,Rossig L,Koehl U,Koyanagi M,Mohamed A,Brandes RP,Martin H,Zeiher AM,Dimmeler S

    更新日期:2006-09-26 00:00:00

  • Formation of IgE-binding factors by T cells of patients infected with human immunodeficiency virus type 1.

    abstract::Peripheral blood mononuclear cells of 10 out of 26 patients with human immunodeficiency virus type 1 (HIV-1) released IgE-binding factors, as determined by two independent assays. The formation of the factors by the mononuclear cells was enhanced by incubation of the cells with homologous IgE. In the presence of IgE, ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.85.23.9214

    authors: Carini C,Margolick J,Yodoi J,Ishizaka K

    更新日期:1988-12-01 00:00:00

  • The RNA helicase Dhx15 mediates Wnt-induced antimicrobial protein expression in Paneth cells.

    abstract::RNA helicases play roles in various essential biological processes such as RNA splicing and editing. Recent in vitro studies show that RNA helicases are involved in immune responses toward viruses, serving as viral RNA sensors or immune signaling adaptors. However, there is still a lack of in vivo data to support the ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.2017432118

    authors: Wang Y,He K,Sheng B,Lei X,Tao W,Zhu X,Wei Z,Fu R,Wang A,Bai S,Zhang Z,Hong N,Ye C,Tian Y,Wang J,Li M,Zhang K,Li L,Yang H,Li HB,Flavell RA,Zhu S

    更新日期:2021-01-26 00:00:00

  • Dysregulated gene expression networks in human acute myelogenous leukemia stem cells.

    abstract::We performed the first genome-wide expression analysis directly comparing the expression profile of highly enriched normal human hematopoietic stem cells (HSC) and leukemic stem cells (LSC) from patients with acute myeloid leukemia (AML). Comparing the expression signature of normal HSC to that of LSC, we identified 3...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0900089106

    authors: Majeti R,Becker MW,Tian Q,Lee TL,Yan X,Liu R,Chiang JH,Hood L,Clarke MF,Weissman IL

    更新日期:2009-03-03 00:00:00