spheroplasts

Summary

Summary: Cells, usually bacteria or yeast, which have partially lost their cell wall, lost their characteristic shape and become round.

Top Publications

  1. Anishkin A, Chiang C, Sukharev S. Gain-of-function mutations reveal expanded intermediate states and a sequential action of two gates in MscL. J Gen Physiol. 2005;125:155-70 pubmed
    ..These data strongly suggest that the bulk of conductance gain in the second subtransition (S-->O) occurs through the opening of the NH2-terminal (S1) gate and the linkers are coupling elements between the M1 and S1 gates...
  2. Akitake B, Anishkin A, Sukharev S. The "dashpot" mechanism of stretch-dependent gating in MscS. J Gen Physiol. 2005;125:143-54 pubmed
    ..Here we characterized the gating of MscS in Escherichia coli spheroplasts in a triple mutant (mscL-, mscS-, mscK-) background...
  3. Tefsen B, Geurtsen J, Beckers F, Tommassen J, de Cock H. Lipopolysaccharide transport to the bacterial outer membrane in spheroplasts. J Biol Chem. 2005;280:4504-9 pubmed
    ..Whereas newly synthesized lipoproteins could be released from spheroplasts of Escherichia coli upon addition of a periplasmic extract containing LolA, de novo synthesized LPS was not ..
  4. King C, Wang H, Chang H. Transformation of yeast by infectious prion particles. Methods. 2006;39:68-71 pubmed
    ..The methodology can be used to introduce different [PSI(+)] particles to many laboratory yeast genetic backgrounds. It can be adapted for applications in other yeast prion systems as well...
  5. Sukharev S, Martinac B, Arshavsky V, Kung C. Two types of mechanosensitive channels in the Escherichia coli cell envelope: solubilization and functional reconstitution. Biophys J. 1993;65:177-83 pubmed
    ..MSCs from Escherichia coli (strain AW740) were examined using the patch-clamp technique, either (a) in giant spheroplasts, (b) after reconstitution by fusing native membrane vesicles with asolectin liposomes, or (c) by reassembly of ..
  6. Hines M, Styer E. Preliminary characterization of chemically generated Mycobacterium avium subsp. paratuberculosis cell wall deficient forms (spheroplasts). Vet Microbiol. 2003;95:247-58 pubmed
    Cell wall deficient forms (CWD, spheroplasts) genetically indistinguishable from M. avium subsp. paratuberculosis (MAP) have been isolated from patients with Crohn's disease and sarcoidosis...
  7. Sakuragi S, Sakuragi J, Morikawa Y, Shioda T. Development of a rapid and convenient method for the quantification of HIV-1 budding. Microbes Infect. 2006;8:1875-81 pubmed
    ..We have previously reported that spheroplasts of Saccharomyces cerevisiae expressing HIV-1 Gag proteins from an episomal plasmid constitutively released a ..
  8. Kwok A, Mak C, Wong F, Wong J. Novel method for preparing spheroplasts from cells with an internal cellulosic cell wall. Eukaryot Cell. 2007;6:563-7 pubmed
  9. Sakuragi S, Goto T, Sano K, Morikawa Y. HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 2002;99:7956-61 pubmed
    ..show that, after removal of the cell wall, the expression of HIV type 1 Gag protein in Saccharomyces cerevisiae spheroplasts allowed simultaneous budding of VLPs from the plasma membrane...

More Information

Publications62

  1. Jung S, Jeong K, Iverson B, Georgiou G. Binding and enrichment of Escherichia coli spheroplasts expressing inner membrane tethered scFv antibodies on surface immobilized antigens. Biotechnol Bioeng. 2007;98:39-47 pubmed
    ..Conversion of the cells into spheroplasts and incubation with soluble fluorescently conjugated ligands results in the specific labeling of cells ..
  2. Noskov V, Kouprina N, Leem S, Koriabine M, Barrett J, Larionov V. A genetic system for direct selection of gene-positive clones during recombinational cloning in yeast. Nucleic Acids Res. 2002;30:E8 pubmed
    ..This method produces positive YAC recombinants at a frequency of approximately 40%. This novel TAR cloning method provides a powerful tool for structural and functional analysis of complex genomes...
  3. Sullivan C, Morrell J, Allison D, Doktycz M. Mounting of Escherichia coli spheroplasts for AFM imaging. Ultramicroscopy. 2005;105:96-102 pubmed
    ..AFM images demonstrate that spheroplasts can be secured with warm gelatin applied to the mica substrate just before the addition of a spheroplast ..
  4. Macdonald A, Martinac B. Effect of high hydrostatic pressure on the bacterial mechanosensitive channel MscS. Eur Biophys J. 2005;34:434-41 pubmed
    ..These observations also have implications for our understanding of the adaptation of mechanosensitive channels in deep-sea bacteria...
  5. Qiu J, Jung S, Georgiou G, Hang H. Enrichment of Escherichia coli spheroplasts displaying scFv antibodies specific for antigens expressed on the human cell surface. Appl Microbiol Biotechnol. 2010;88:1385-91 pubmed publisher
    ..In this study, we demonstrate that Escherichia coli spheroplasts, displaying antibodies and screened by a combination of plate-panning and flow cytometric sorting, can be used ..
  6. Kamaraju K, Gottlieb P, Sachs F, Sukharev S. Effects of GsMTx4 on bacterial mechanosensitive channels in inside-out patches from giant spheroplasts. Biophys J. 2010;99:2870-8 pubmed publisher
    ..the effects of cytoplasmic GsMTx4 on the two bacterial channels, MscS and MscL, in giant Escherichia coli spheroplasts. In excised inside-out patches, GsMTx4 sensitized both channels to tension by increasing the opening rate and ..
  7. Yoshimura K, Batiza A, Schroeder M, Blount P, Kung C. Hydrophilicity of a single residue within MscL correlates with increased channel mechanosensitivity. Biophys J. 1999;77:1960-72 pubmed
    ..These results suggest a model for gating in which Gly(22) moves from a hydrophobic, and through a hydrophilic, environment upon transition from the closed to open conformation...
  8. Kelly F, Nurse P. De novo growth zone formation from fission yeast spheroplasts. PLoS ONE. 2011;6:e27977 pubmed publisher
    ..As spheroplasts recover, new growth zones form that resemble normal growing cell tips in shape and width, and polarized growth ..
  9. Nakamaru Y, Takahashi Y, Unemoto T, Nakamura T. Mechanosensitive channel functions to alleviate the cell lysis of marine bacterium, Vibrio alginolyticus, by osmotic downshock. FEBS Lett. 1999;444:170-2 pubmed
    ..This is the first report to show a physiological role of the mechanosensitive channel with large conductance...
  10. Moe P, Blount P, Kung C. Functional and structural conservation in the mechanosensitive channel MscL implicates elements crucial for mechanosensation. Mol Microbiol. 1998;28:583-92 pubmed
    ..This structural conservation is providing insight into regions of this molecule that are vital to its role as a mechanosensitive channel and may have broader implications for the understanding of other mechanosensitive systems...
  11. Hurst A, Gottlieb P, Martinac B. Concentration dependent effect of GsMTx4 on mechanosensitive channels of small conductance in E. coli spheroplasts. Eur Biophys J. 2009;38:415-25 pubmed publisher
    ..coli spheroplasts using the patch-clamp technique...
  12. Sukharev S, Blount P, Martinac B, Blattner F, Kung C. A large-conductance mechanosensitive channel in E. coli encoded by mscL alone. Nature. 1994;368:265-8 pubmed
    ..The mscL nucleotide sequence predicts a unique protein of only 136 amino acids, with a highly hydrophobic core and very different from porins or other known proteins...
  13. Nomura T, Cranfield C, Deplazes E, Owen D, Macmillan A, Battle A, et al. Differential effects of lipids and lyso-lipids on the mechanosensitivity of the mechanosensitive channels MscL and MscS. Proc Natl Acad Sci U S A. 2012;109:8770-5 pubmed publisher
  14. Narita S, Tanaka K, Matsuyama S, Tokuda H. Disruption of lolCDE, encoding an ATP-binding cassette transporter, is lethal for Escherichia coli and prevents release of lipoproteins from the inner membrane. J Bacteriol. 2002;184:1417-22 pubmed
    ..From these results, we conclude that the LolCDE complex is an essential ABC transporter for E. coli and the sole apparatus mediating the release of outer membrane lipoproteins from the inner membrane...
  15. Sanchez C, Preuss M, Lanzer M. Construction and screening of YAC libraries. Methods Mol Med. 2002;72:291-304 pubmed
  16. Bienert G, Møller A, Kristiansen K, Schulz A, Møller I, Schjoerring J, et al. Specific aquaporins facilitate the diffusion of hydrogen peroxide across membranes. J Biol Chem. 2007;282:1183-92 pubmed
    ..Our results present the first molecular genetic evidence for the diffusion of H2O2 through specific members of the aquaporin family...
  17. Elvira Recuenco M, van Vuurde J. Efficiency of procedures for induction and cultivation of Pseudomonas syringae pv. pisi L-form. Microbiol Res. 2003;158:271-9 pubmed
    ..Based on the results from these tests an induction method for P. s. pv. pisi L-form is proposed and the relevance of L-form is discussed for practice...
  18. Thomas D, Bron P, Weimann T, Dautant A, Giraud M, Paumard P, et al. Supramolecular organization of the yeast F1Fo-ATP synthase. Biol Cell. 2008;100:591-601 pubmed publisher
    ..For a large range of organisms, it has been shown that mitochondrial ATP synthase adopts oligomeric structures. Moreover, several studies have suggested that a link exists between ATP synthase and mitochondrial morphology...
  19. Belyy V, Kamaraju K, Akitake B, Anishkin A, Sukharev S. Adaptive behavior of bacterial mechanosensitive channels is coupled to membrane mechanics. J Gen Physiol. 2010;135:641-52 pubmed publisher
    ..MscS activities tried in spheroplast-attached mode showed no adaptation when the spheroplasts were intact, but permeabilized spheroplasts showed delayed adaptation, suggesting that the presence of membrane ..
  20. Komiyama T, Kimura T, Furuichi Y. Round shape enlargement of the yeast spheroplast of Saccharomyces cerevisiae by HM-1 toxin. Biol Pharm Bull. 2002;25:959-65 pubmed
    The effects of HM-1 killer toxin (HM-1) on yeast spheroplasts of Saccharomyces cerevisiae were examined under osmotically stabilized conditions...
  21. Mazor Y, Van Blarcom T, Carroll S, Georgiou G. Selection of full-length IgGs by tandem display on filamentous phage particles and Escherichia coli fluorescence-activated cell sorting screening. FEBS J. 2010;277:2291-303 pubmed publisher
  22. Masi M, Duret G, Delcour A, Misra R. Folding and trimerization of signal sequence-less mature TolC in the cytoplasm of Escherichia coli. Microbiology. 2009;155:1847-57 pubmed publisher
    ..Genetic and biochemical data attribute the unique folding and assembly pathways of TolC to its large soluble alpha-helical domain...
  23. Baggett J, Shaw J, Sciambi C, Watson H, Wendland B. Fluorescent labeling of yeast. Curr Protoc Cell Biol. 2003;Chapter 4:Unit 4.13 pubmed publisher
  24. Graham J. Isolation of peroxisomes from tissues and cells by differential and density gradient centrifugation. Curr Protoc Cell Biol. 2001;Chapter 3:Unit 3.5 pubmed publisher
    ..unit contains protocols for peroxisome isolation from rat liver, tissue culture cells (HepG2 cells), and yeast spheroplasts. The extent of endoplasmic reticulum contamination of the prep can be assessed using an assay for the marker ..
  25. Cavalheiro R, Fortes F, Borecky J, Faustinoni V, Schreiber A, Vercesi A. Respiration, oxidative phosphorylation, and uncoupling protein in Candida albicans. Braz J Med Biol Res. 2004;37:1455-61 pubmed
    ..membrane potential (Deltapsi), and oxidative phosphorylation of mitochondria in situ were determined in spheroplasts obtained from Candida albicans control strain ATCC 90028 by lyticase treatment...
  26. Codling C, Maillard J, Russell A. Aspects of the antimicrobial mechanisms of action of a polyquaternium and an amidoamine. J Antimicrob Chemother. 2003;51:1153-8 pubmed
    ..These were Acanthamoeba castellanii (trophozoites and cysts), Aspergillus fumigatus, Candida albicans, Pseudomonas aeruginosa, Serratia marcescens and Staphylococcus aureus...
  27. Smarda J, Matejková P, Vavrícková A. Translocation of colicin from the receptor to the inner cell membrane: function of the peptidoglycan layer. Folia Microbiol (Praha). 2002;47:213-7 pubmed
    Sensitivity of spheroplasts (prepared in two ways) of a colicin-sensitive strain, of colicin-resistant and of colicin-tolerant mutants and of strains immune to colicins E1 and E2 was estimated and compared...
  28. Lin J, Liao T, Lee H, King C. Inter-allelic prion propagation reveals conformational relationships among a multitude of [PSI] strains. PLoS Genet. 2011;7:e1002297 pubmed publisher
    ..It thus suggests a possible general scheme to classify a multitude of prion strains...
  29. Ramanandraibe E, Younsi M, Coulon J, Loppinet V, Hakkou A, Bonaly R. Implication of cell wall constituents in the sensitivity of Kluyveromyces lactis strains to amphotericin B. Res Microbiol. 1998;149:109-18 pubmed
    ..The present results reveal that both a change in the ionic charges of the cell wall and an alteration in the cell wall structure modified the sensitivity of these yeast strains to AmB...
  30. Rowe I, Elahi M, Huq A, Sukharev S. The mechanoelectrical response of the cytoplasmic membrane of Vibrio cholerae. J Gen Physiol. 2013;142:75-85 pubmed publisher
    ..Direct electrophysiology of V. cholerae offers new avenues for the in situ analysis of membrane components critical for osmotic survival and electrogenic transport in this pathogen. ..
  31. Sevova E, Goren M, Schwartz K, Hsu F, Turk J, Fox B, et al. Cell-free synthesis and functional characterization of sphingolipid synthases from parasitic trypanosomatid protozoa. J Biol Chem. 2010;285:20580-7 pubmed publisher
    ..These results represent the first application of cell-free synthesis for the rapid preparation and functional annotation of integral membrane proteins and thus illustrate its utility in studying otherwise intractable enzyme systems...
  32. Mann B, Bowman B, Grotelueschen J, Metzenberg R. Nucleotide sequence of pho-4+, encoding a phosphate-repressible phosphate permease of Neurospora crassa. Gene. 1989;83:281-9 pubmed
    ..Since very few permease-encoding genes of eukaryotes have been examined in molecular detail, it will be of interest to compare the sequence of pho-4+ with those encoding other anion transport proteins, as they become available...
  33. Schones D, Cui K, Cuddapah S. Genome-wide approaches to studying yeast chromatin modifications. Methods Mol Biol. 2011;759:61-71 pubmed publisher
  34. Ma L, Narberhaus F, Lai E. IcmF family protein TssM exhibits ATPase activity and energizes type VI secretion. J Biol Chem. 2012;287:15610-21 pubmed publisher
    ..Our findings strongly argue that TssM functions as a T6SS energizer to recruit Hcp into the TssM-TssL inner membrane complex prior to Hcp secretion across the outer membrane...
  35. Wegner A, Alexeeva S, Odijk T, Woldringh C. Characterization of Escherichia coli nucleoids released by osmotic shock. J Struct Biol. 2012;178:260-9 pubmed publisher
    Nucleoids were isolated by osmotic shock from Escherichia coli spheroplasts at relatively low salt concentrations and in the absence of detergents...
  36. Ranjit D, Young K. The Rcs stress response and accessory envelope proteins are required for de novo generation of cell shape in Escherichia coli. J Bacteriol. 2013;195:2452-62 pubmed publisher
    ..Though the spheroplasts thus created usually lyse, some may survive by resynthesizing their walls and shapes...
  37. Tao K. Subcellular localization and in vivo oxidation-reduction kinetics of thiol peroxidase in Escherichia coli. FEMS Microbiol Lett. 2008;289:41-5 pubmed publisher
    ..Reanalysis of Tpx localization, using active site cysteine mutants of thioredoxin 1 (Trx1), demonstrated that Tpx forms a mixed-disulfide complex with cytoplasmic Trx1, indicating that Tpx localizes in the cytoplasm...
  38. Haarasilta S, Taskinen L. Location of three key enzymes of gluconeogenesis in baker's yeast. Arch Microbiol. 1977;113:159-61 pubmed
    ..investigated by density gradient centrifugation of spheroplast lysates obtained by osmotic shock treatment of spheroplasts and centrifugation for 10000 g x min...
  39. Friedberg I. Localization of phosphoglucose isomerase in Escherichia coli and its relation to the induction of the hexose phosphate transport system. J Bacteriol. 1972;112:1201-5 pubmed
    ..S6P was not taken by intact cells. The data indicate that the isomerization of F6P to G6P can take place on the cell surface, and this explains the indirect induction of hexose phosphate transport by F6P...
  40. Zimmann P, Puppe W, Altendorf K. Membrane topology analysis of the sensor kinase KdpD of Escherichia coli. J Biol Chem. 1995;270:28282-8 pubmed
    ..was investigated using LacZ and PhoA fusions at different sites within the polypeptide chain and by treating spheroplasts in the presence or absence of Triton X-100 with the protease kallikrein...
  41. Zhou J, Fazzio R, Blair D. Membrane topology of the MotA protein of Escherichia coli. J Mol Biol. 1995;251:237-42 pubmed
    ..These experiments establish that the main features of the suggested model for MotA topology are correct, furnishing a basis for more detailed structure-function studies of the MotA/MotB proton channel...
  42. Plesofsky Vig N, Brambl R. Disruption of the gene for hsp30, an alpha-crystallin-related heat shock protein of Neurospora crassa, causes defects in thermotolerance. Proc Natl Acad Sci U S A. 1995;92:5032-6 pubmed
    ..Results of this study suggest that hsp30 may be important for efficient carbohydrate utilization during high temperature stress and that it may interact with other mitochondrial membrane proteins and function as a protein chaperone...
  43. Vida T, Gerhardt B. A cell-free assay allows reconstitution of Vps33p-dependent transport to the yeast vacuole/lysosome. J Cell Biol. 1999;146:85-98 pubmed
    We report a cell-free system that measures transport-coupled maturation of carboxypeptidase Y (CPY). Yeast spheroplasts are lysed by extrusion through polycarbonate filters...
  44. Kosova B, Pante N, Rollenhagen C, Podtelejnikov A, Mann M, Aebi U, et al. Mlp2p, a component of nuclear pore attached intranuclear filaments, associates with nic96p. J Biol Chem. 2000;275:343-50 pubmed
    ..When Nic96p was affinity purified from glutaraldehyde-treated spheroplasts, it was found to be associated with Mlp2p...
  45. Zhao R, Reithmeier R. Expression and characterization of the anion transporter homologue YNL275w in Saccharomyces cerevisiae. Am J Physiol Cell Physiol. 2001;281:C33-45 pubmed
    ..These results suggest that the yeast gene YNL275w encodes a nonglycosylated anion transport protein, localized to the plasma membrane...
  46. Kaufman B, Newman S, Perlman P, Butow R. Crosslinking of proteins to mtDNA. Methods Mol Biol. 2002;197:377-89 pubmed
  47. Rodal A, Kozubowski L, Goode B, Drubin D, Hartwig J. Actin and septin ultrastructures at the budding yeast cell cortex. Mol Biol Cell. 2005;16:372-84 pubmed
    ..These images provide the first high-resolution views of septin organization in cells...
  48. Melent ev A, Helisto P, Kuz mina L, Galimzianova N, Aktuganov G, Korpela T. [Application of Bacillus-antagonists for biocontrol of fungi degrading raw wood]. Prikl Biokhim Mikrobiol. 2006;42:70-5 pubmed
    ..antagonistic bacilli inhibited growth and development of wood-decay fungi in vitro and led to the formation of spheroplasts on growing hyphae...
  49. Meluh P, Broach J. Immunological analysis of yeast chromatin. Methods Enzymol. 1999;304:414-30 pubmed
  50. King S, Kenney L. Application of fluorescence resonance energy transfer to examine EnvZ/OmpR interactions. Methods Enzymol. 2007;422:352-60 pubmed
    ..b>Spheroplasts were prepared and lysed in microtiter plates containing purified, fluorescent-labeled OmpR protein...
  51. Foote S, Denny C. Construction of YAC libraries with large inserts. Curr Protoc Hum Genet. 2002;Chapter 5:Unit5.2 pubmed publisher
    ..basic protocol in this unit describes preparation of YAC vectors and transformation of ligated DNA into yeast spheroplasts. A support protocol describes titration of Lyticase to make spheroplasts...
  52. Wang Y, Liao M, Hoe N, Acharya P, Deng C, Krutchinsky A, et al. A role for protein phosphorylation in cytochrome P450 3A4 ubiquitin-dependent proteasomal degradation. J Biol Chem. 2009;284:5671-84 pubmed publisher
  53. Ferrante T, Viola F, Balliano G, Oliaro Bosso S. Difference in the late ergosterol biosynthesis between yeast spheroplasts and intact cells. Acta Biochim Pol. 2016;63:371-5 pubmed publisher
    A comparative study on post-squalene sterol synthesis in intact yeast cells and spheroplasts was carried out with strains from three genera (Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris) as well as with engineered ..