Real-time attack on single Escherichia coli cells by the human antimicrobial peptide LL-37.

Abstract:

:Natural antimicrobial peptides (AMPs) provide prototypes for the design of unconventional antimicrobial agents. Existing bulk assays measure AMP activity but do not provide details of the growth-halting mechanism. We use fluorescence microscopy to directly observe the attack of the human antimicrobial peptide LL-37 on single Escherichia coli cells in real time. Our findings strongly suggest that disruption of the cytoplasmic membrane is not the growth-halting mechanism. At 8 μM, LL-37 binding saturates the outer membrane (OM) within 1 min. Translocation across the OM and access to the periplasmic space (5-25 min later) correlates in time with the halting of growth. Septating cells are attacked more readily than nonseptating cells. The halting of growth may occur because of LL-37 interference with cell wall biogenesis. Only well after growth halts does the peptide permeabilize the cytoplasmic membrane to GFP and the small dye Sytox Green. The assay enables dissection of antimicrobial design criteria into two parts: translocation across the OM and the subsequent halting of growth.

authors

Sochacki KA,Barns KJ,Bucki R,Weisshaar JC

doi

10.1073/pnas.1101130108

subject

Has Abstract

pub_date

2011-04-19 00:00:00

pages

E77-81

issue

16

eissn

0027-8424

issn

1091-6490

pii

1101130108

journal_volume

108

pub_type

杂志文章
  • Enantiomer discrimination illustrated by high-resolution crystal structures of the human nuclear receptor hRARgamma.

    abstract::The human retinoic acid receptor (hRAR) is a member of the nuclear receptor superfamily that regulates the transcription of target genes in a ligand-dependent manner. The three hRAR isotypes are targets for retinoids that are used in the treatment of various diseases, including breast cancer and skin diseases. Drug ef...

    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.97.12.6322

    authors: Klaholz BP,Mitschler A,Belema M,Zusi C,Moras D

    更新日期:2000-06-06 00:00:00

  • Multiple rare variants in different genes account for multifactorial inherited susceptibility to colorectal adenomas.

    abstract::Clear-cut inherited Mendelian traits, such as familial adenomatous polyposis or hereditary nonpolyposis colorectal cancer, account for <4% of colorectal cancers. Another 20% of all colorectal cancers are thought to occur in individuals with a significant inherited multifactorial susceptibility to colorectal cancer tha...

    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.0407187101

    authors: Fearnhead NS,Wilding JL,Winney B,Tonks S,Bartlett S,Bicknell DC,Tomlinson IP,Mortensen NJ,Bodmer WF

    更新日期:2004-11-09 00:00:00

  • Chemical composition of Earth, Venus, and Mercury.

    abstract::Model compositions of Earth, Venus, and Mercury are calculated from the premise that planets and chondrites underwent four identical fractionation processes in the solar nebula. Because elements of similar properties stay together in these processes, five constraints suffice to define the composition of a planet: mass...

    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.77.12.6973

    authors: Morgan JW,Anders E

    更新日期:1980-12-01 00:00:00

  • Akt-dependent phosphorylation of endothelial nitric-oxide synthase mediates penile erection.

    abstract::In the penis, nitric oxide (NO) can be formed by both neuronal NO synthase and endothelial NOS (eNOS). eNOS is activated by viscous drag/shear stress in blood vessels to produce NO continuously, a process mediated by the phosphatidylinositol 3-kinase (PI3kinase)/Akt pathway. Here we show that PI3-kinase/Akt physiologi...

    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.052712499

    authors: Hurt KJ,Musicki B,Palese MA,Crone JK,Becker RE,Moriarity JL,Snyder SH,Burnett AL

    更新日期:2002-03-19 00:00:00

  • Heme oxygenase induction mediates the photoimmunoprotective activity of UVA radiation in the mouse.

    abstract::In contrast to the immunosuppressive potential of UVB (280-320 nm) radiation in experimental animals and humans, UVA (320-400 nm) radiation at environmentally relevant doses appears to be immunologically inert. However, such exposure to UVA radiation has been observed unexpectedly to induce resistance to UVB-induced i...

    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.96.16.9317

    authors: Reeve VE,Tyrrell RM

    更新日期:1999-08-03 00:00:00

  • O-GlcNAc reports ambient temperature and confers heat resistance on ectotherm development.

    abstract::Effects of temperature on biological processes are complex. Diffusion is less affected than the diverse enzymatic reactions that have distinct individual temperature profiles. Hence thermal fluctuations pose a formidable challenge to ectothermic organisms in which body temperature is largely dictated by the ambient te...

    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.1322396111

    authors: Radermacher PT,Myachina F,Bosshardt F,Pandey R,Mariappa D,Müller HA,Lehner CF

    更新日期:2014-04-15 00:00:00

  • Shoot-and-Trap: use of specific x-ray damage to study structural protein dynamics by temperature-controlled cryo-crystallography.

    abstract::Although x-ray crystallography is the most widely used method for macromolecular structure determination, it does not provide dynamical information, and either experimental tricks or complementary experiments must be used to overcome the inherently static nature of crystallographic structures. Here we used specific x-...

    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.0804828105

    authors: Colletier JP,Bourgeois D,Sanson B,Fournier D,Sussman JL,Silman I,Weik M

    更新日期:2008-08-19 00:00:00

  • Endothelial microRNA-150 is an intrinsic suppressor of pathologic ocular neovascularization.

    abstract::Pathologic ocular neovascularization commonly causes blindness. It is critical to identify the factors altered in pathologically proliferating versus normally quiescent vessels to develop effective targeted therapeutics. MicroRNAs regulate both physiological and pathological angiogenesis through modulating expression ...

    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.1508426112

    authors: Liu CH,Sun Y,Li J,Gong Y,Tian KT,Evans LP,Morss PC,Fredrick TW,Saba NJ,Chen J

    更新日期:2015-09-29 00:00:00

  • The Vibrio cholerae O139 O-antigen polysaccharide is essential for Ca2+-dependent biofilm development in sea water.

    abstract::Vibrio cholerae is both an inhabitant of estuarine environments and the etiologic agent of the diarrheal disease cholera. Previous work has demonstrated that V. cholerae forms both an exopolysaccharide-dependent biofilm and a Ca2+-dependent biofilm. In this work, we demonstrate a role for the O-antigen polysaccharide ...

    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.2334614100

    authors: Kierek K,Watnick PI

    更新日期:2003-11-25 00:00:00

  • Serotonin 2B receptor is required for heart development.

    abstract::Several lines of evidence suggest that the serotonin (5-hydroxytryptamine, 5-HT) regulates cardiovascular functions during embryogenesis and adulthood. 5-HT binds to numerous cognate receptors to initiate its biological effects. However, none of the 5-HT receptor disruptions in mice have yet resulted in embryonic defe...

    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.97.17.9508

    authors: Nebigil CG,Choi DS,Dierich A,Hickel P,Le Meur M,Messaddeq N,Launay JM,Maroteaux L

    更新日期:2000-08-15 00:00:00

  • Structural basis for antibody recognition of the NANP repeats in Plasmodium falciparum circumsporozoite protein.

    abstract::Acquired resistance against antimalarial drugs has further increased the need for an effective malaria vaccine. The current leading candidate, RTS,S, is a recombinant circumsporozoite protein (CSP)-based vaccine against Plasmodium falciparum that contains 19 NANP repeats followed by a thrombospondin repeat domain. Alt...

    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.1715812114

    authors: Oyen D,Torres JL,Wille-Reece U,Ockenhouse CF,Emerling D,Glanville J,Volkmuth W,Flores-Garcia Y,Zavala F,Ward AB,King CR,Wilson IA

    更新日期:2017-11-28 00:00:00

  • Viable hypomorphic signaling mutant of the Met receptor reveals a role for hepatocyte growth factor in postnatal cerebellar development.

    abstract::Cerebellar development occurs mainly postnatally and implies cell proliferation and migration. Hepatocyte growth factor (HGF) and Met are involved in mediating these responses in other tissues and are coexpressed in the cerebellum. Here we show that Met is localized in granule cell precursors and that cultures of thes...

    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.222362099

    authors: Ieraci A,Forni PE,Ponzetto C

    更新日期:2002-11-12 00:00:00

  • The FLP recombinase of the yeast 2-micron plasmid: characterization of its recombination site.

    abstract::The minimal size of the recombination site required for efficient FLP recombinase-catalyzed recombination in vitro is no more than 28 base pairs, which includes parts of two 13-base-pair inverted repeats and all of an 8-base-pair spacer. The FLP recombinase cleaves the DNA at the boundaries of the spacer, becomes cova...

    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.82.21.7270

    authors: Senecoff JF,Bruckner RC,Cox MM

    更新日期:1985-11-01 00:00:00

  • DNase I-hypersensitive sites in the chromatin of immunoglobulin kappa light chain genes.

    abstract::We have used DNase I as a probe to examine the chromatin structure of mouse immunoglobulin kappa light chain genes in rearranged and unrearranged chromosomes--i.e., in nuclei from myeloma cells and from brain and liver cells. Tissue-specific DNase I-hypersensitive sites are observed 0.7 and 1.7 kilobases upstream from...

    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.80.9.2427

    authors: Chung SY,Folsom V,Wooley J

    更新日期:1983-05-01 00:00:00

  • Epidermal growth factor stimulates tyrosine phosphorylation in the neonatal mouse: association of a M(r) 55,000 substrate with the receptor.

    abstract::Administration of epidermal growth factor (EGF) to neonatal mice resulted in rapid tyrosine phosphorylation of a number of specific substrates in liver, kidney, lung, bladder, skin, and brain as detected by Western blot analysis of tissue homogenates with anti-phosphotyrosine antibodies. In the liver, three prominent ...

    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.89.18.8477

    authors: Donaldson RW,Cohen S

    更新日期:1992-09-15 00:00:00

  • T lymphocytes that infiltrate tumors and atherosclerotic plaques produce heparin-binding epidermal growth factor-like growth factor and basic fibroblast growth factor: a potential pathologic role.

    abstract::Despite significant infiltration into tumors and atherosclerotic plaques, the role of T lymphocytes in these pathological conditions is still unclear. We have demonstrated that tumor-infiltrating lymphocytes (TILs) and plaque-infiltrating lymphocytes (PILs) produce heparin-binding epidermal growth factor-like growth f...

    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.92.14.6547

    authors: Peoples GE,Blotnick S,Takahashi K,Freeman MR,Klagsbrun M,Eberlein TJ

    更新日期:1995-07-03 00:00:00

  • Mouse lipin-1 and lipin-2 cooperate to maintain glycerolipid homeostasis in liver and aging cerebellum.

    abstract::The three lipin phosphatidate phosphatase (PAP) enzymes catalyze a step in glycerolipid biosynthesis, the conversion of phosphatidate to diacylglycerol. Lipin-1 is critical for lipid synthesis and homeostasis in adipose tissue, liver, muscle, and peripheral nerves. Little is known about the physiological role of lipin...

    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.1205221109

    authors: Dwyer JR,Donkor J,Zhang P,Csaki LS,Vergnes L,Lee JM,Dewald J,Brindley DN,Atti E,Tetradis S,Yoshinaga Y,De Jong PJ,Fong LG,Young SG,Reue K

    更新日期:2012-09-11 00:00:00

  • Calcium waves occur as Drosophila oocytes activate.

    abstract::Egg activation is the process by which a mature oocyte becomes capable of supporting embryo development. In vertebrates and echinoderms, activation is induced by fertilization. Molecules introduced into the egg by the sperm trigger progressive release of intracellular calcium stores in the oocyte. Calcium wave(s) spre...

    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.1420589112

    authors: Kaneuchi T,Sartain CV,Takeo S,Horner VL,Buehner NA,Aigaki T,Wolfner MF

    更新日期:2015-01-20 00:00:00

  • Amplification of DNA from preserved specimens shows blowflies were preadapted for the rapid evolution of insecticide resistance.

    abstract::Mutations of esterase 3 confer two forms of organophosphate resistance on contemporary Australasian Lucilia cuprina. One form, called diazinon resistance, is slightly more effective against commonly used insecticides and is now more prevalent than the other form, called malathion resistance. We report here that the si...

    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.0509590103

    authors: Hartley CJ,Newcomb RD,Russell RJ,Yong CG,Stevens JR,Yeates DK,La Salle J,Oakeshott JG

    更新日期:2006-06-06 00:00:00

  • Identification of a second aryl phosphate-binding site in protein-tyrosine phosphatase 1B: a paradigm for inhibitor design.

    abstract::The structure of the catalytically inactive mutant (C215S) of the human protein-tyrosine phosphatase 1B (PTP1B) has been solved to high resolution in two complexes. In the first, crystals were grown in the presence of bis-(para-phosphophenyl) methane (BPPM), a synthetic high-affinity low-molecular weight nonpeptidic s...

    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.94.25.13420

    authors: Puius YA,Zhao Y,Sullivan M,Lawrence DS,Almo SC,Zhang ZY

    更新日期:1997-12-09 00:00:00

  • Broadly neutralizing anti-HIV-1 antibodies disrupt a hinge-related function of gp41 at the membrane interface.

    abstract::A vaccine capable of stimulating protective antiviral antibody responses is needed to curtail the global AIDS epidemic caused by HIV-1. Although rarely elicited during the course of natural infection or upon conventional vaccination, the membrane-proximal ectodomain region (MPER) of the HIV-1 glycoprotein of M(r) 41,0...

    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.0901474106

    authors: Song L,Sun ZY,Coleman KE,Zwick MB,Gach JS,Wang JH,Reinherz EL,Wagner G,Kim M

    更新日期:2009-06-02 00:00:00

  • An artificial cell-cycle inhibitor isolated from a combinatorial library.

    abstract::Understanding the genetic networks that operate inside cells will require the dissection of interactions among network members. Here we describe a peptide aptamer isolated from a combinatorial library that distinguishes among such interactions. This aptamer binds to cyclin-dependent kinase 2 (Cdk2) and inhibits its ki...

    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.95.24.14272

    authors: Cohen BA,Colas P,Brent R

    更新日期:1998-11-24 00:00:00

  • The human immunodeficiency virus type 2 vpr gene is essential for productive infection of human macrophages.

    abstract::The human immunodeficiency virus (HIV) genetic determinant(s) responsible for tropism in human T cells or macrophages are not well defined. We studied the role of the HIV type 2 (HIV-2) nef and vpr genes in viral tropism. HIV-2 mutants, lacking either vpr or nef genes, or both vpr and nef, were obtained by site-specif...

    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.87.20.8080

    authors: Hattori N,Michaels F,Fargnoli K,Marcon L,Gallo RC,Franchini G

    更新日期:1990-10-01 00:00:00

  • Stability of plant immune-receptor resistance proteins is controlled by SKP1-CULLIN1-F-box (SCF)-mediated protein degradation.

    abstract::The nucleotide-binding domain and leucine-rich repeats containing proteins (NLRs) serve as immune receptors in both plants and animals. Overaccumulation of NLRs often leads to autoimmune responses, suggesting that the levels of these immune receptors must be tightly controlled. However, the mechanism by which NLR prot...

    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.1105685108

    authors: Cheng YT,Li Y,Huang S,Huang Y,Dong X,Zhang Y,Li X

    更新日期:2011-08-30 00:00:00

  • Glucose sensing by MondoA:Mlx complexes: a role for hexokinases and direct regulation of thioredoxin-interacting protein expression.

    abstract::Glucose is a fundamental metabolite, yet how cells sense and respond to changes in extracellular glucose concentration is not completely understood. We recently reported that the MondoA:Mlx dimeric transcription factor directly regulates glycolysis. In this article, we consider whether MondoA:Mlx complexes have a broa...

    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.0712199105

    authors: Stoltzman CA,Peterson CW,Breen KT,Muoio DM,Billin AN,Ayer DE

    更新日期:2008-05-13 00:00:00

  • A pathogen-inducible endogenous siRNA in plant immunity.

    abstract::RNA interference, mediated by small interfering RNAs (siRNAs), is a conserved regulatory process that has evolved as an antiviral defense mechanism in plants and animals. It is not known whether host cells also use siRNAs as an antibacterial defense mechanism in eukaryotes. Here, we report the discovery of an endogeno...

    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.0608258103

    authors: Katiyar-Agarwal S,Morgan R,Dahlbeck D,Borsani O,Villegas A Jr,Zhu JK,Staskawicz BJ,Jin H

    更新日期:2006-11-21 00:00:00

  • The primary step of biotin synthesis in mycobacteria.

    abstract::Biotin plays an essential role in growth of mycobacteria. Synthesis of the cofactor is essential for Mycobacterium tuberculosis to establish and maintain chronic infections in a murine model of tuberculosis. Although the late steps of mycobacterial biotin synthesis, assembly of the heterocyclic rings, are thought to f...

    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.2010189117

    authors: Hu Z,Cronan JE

    更新日期:2020-09-22 00:00:00

  • Squid Pax-6 and eye development.

    abstract::Pax-6 in vertebrates and its homolog eyeless in Drosophila are known to be essential for eye development. Here we investigate the role of Pax-6 in eye development in another major systematic group, molluscs. We demonstrate that alternatively spliced RNAs derived from a single Pax-6 gene in the squid (Loligo opalescens...

    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.94.6.2421

    authors: Tomarev SI,Callaerts P,Kos L,Zinovieva R,Halder G,Gehring W,Piatigorsky J

    更新日期:1997-03-18 00:00:00

  • Expression and nuclear envelope localization of biologically active fusion glycoprotein gB of herpes simplex virus in mammalian cells using cloned DNA.

    abstract::Herpes simplex virus (HSV) is known to bud from the inner membrane of the nuclear envelope. The structural gene for the glycoprotein gB, which is essential for virus entry and cell fusion induced by HSV type 1, has been cloned in a transient expression vector containing the adenovirus major late promoter, tripartite l...

    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.84.16.5675

    authors: Ali MA,Butcher M,Ghosh HP

    更新日期:1987-08-01 00:00:00

  • Bohr's 1913 molecular model revisited.

    abstract::It is generally believed that the old quantum theory, as presented by Niels Bohr in 1913, fails when applied to few electron systems, such as the H(2) molecule. Here, we find previously undescribed solutions within the Bohr theory that describe the potential energy curve for the lowest singlet and triplet states of 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.0505778102

    authors: Svidzinsky AA,Scully MO,Herschbach DR

    更新日期:2005-08-23 00:00:00