Biosynthetic incorporation of fluorescent carbazolylundecanoic acid into membrane phospholipids of LM cells and determination of quenching constants and partition coefficients of hydrophobic quenchers.

Abstract:

:A fluorescence quenching method was developed for determining partition coefficients and diffusional rates of small molecules in cell membranes. This method involves quenching the fluorescence of carbazole-labeled membranes by hydrophobic molecules that partition into membranes. Cell membrane phospholipids of mouse LM cells in tissue culture were biosynthetically labeled with the carbazole moiety by supplementing the growth media with 11-(9-carbazolyl)undecanoic acid. Plasma membranes, microsomes, and mitochondria were isolated free of nonmembranous neutral lipids, and the incorporation of the fluorescent probe was characterized. Quenching studies of the carbazole moiety by a series of N-substituted picolinium perchlorate salts showed that the carbazole moiety was located in the hydrophobic interior of the membrane bilayer. The carbazole fluorescence also was quenched by the hydrophobic quenchers lindane, methoxychlor, and 1,1-dichloro-2,2-bis(rho-chlorophenyl)ethylene, indicating that these compounds partitioned into the membrane. Stern-Volmer quenching constants determined by fluorescence lifetime and intensity measurements were identical, as expected for dynamic quenching. The effects of different lipid compositions on quenching constants and partition coefficients were determined by comparing different membrane fractions. These parameters also were measured in membranes from cells in which the phospholipid composition was altered by substituting ethanolamine for choline in the growth medium. Changes in the lipid composition produced changes in the bimolecular quenching constants. For example, bimolecular quenching constants for 1,1-dichloro-2,2-bis(rho-chlorophenyl)ethylene were higher in mitochondrial membranes than in plasma membranes and microsomes. They were also higher in dispersions made from membrane phospholipids as compared with intact membranes or total lipid dispersion.(ABSTRACT TRUNCATED AT 250 WORDS)

journal_name

Biochemistry

journal_title

Biochemistry

authors

Omann GM,Glaser M

doi

10.1021/bi00316a021

subject

Has Abstract

pub_date

1984-10-09 00:00:00

pages

4962-9

issue

21

eissn

0006-2960

issn

1520-4995

journal_volume

23

pub_type

杂志文章
  • Phosphorylation of beta III-tubulin.

    abstract::There is considerable evidence that mammalian beta-tubulin is phosphorylated. Specifically, of the seven beta isotypes, the phosphorylated one is beta III, the isotype found almost entirely in neurons. The phosphate is added at a serine and perhaps a tyrosine near the C-terminus. All the evidence to date has been gath...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi951247p

    authors: Khan IA,Ludueña RF

    更新日期:1996-03-26 00:00:00

  • "Dilysine trigger" in transferrins probed by mutagenesis of lactoferrin: crystal structures of the R210G, R210E, and R210L mutants of human lactoferrin.

    abstract::The mammalian iron-binding proteins lactoferrin (Lf) and transferrin (Tf) bind iron very tightly, but reversibly. Despite homologous structures and essentially identical iron binding sites, Tf begins to release iron at pH 6.0, whereas Lf retains iron to pH approximately 3.5. This difference in iron retention gives the...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi020443a

    authors: Peterson NA,Arcus VL,Anderson BF,Tweedie JW,Jameson GB,Baker EN

    更新日期:2002-12-03 00:00:00

  • Cardiac troponin T forms a tetramer in vitro.

    abstract::Cardiac troponin T (cTnT) is a myofibrillar protein essential for calcium-dependent contraction. This property has led to functional studies of developmentally expressed cTnT isoforms and mutants identified in patients with hypertrophic cardiomyopathy. The release of cTnT into the serum following myocardial infarction...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi7012596

    authors: Lounes KC,Demeler B,Anderson DE,Gomes AW,Potter JD,Nassar R,Anderson PA

    更新日期:2008-02-19 00:00:00

  • Conformational transition required for efficient splicing of transcripts from hybrid lambda promoter yeast tRNA gene fusion.

    abstract::Fusion of a prokaryotic promoter to a yeast tRNA gene provides a means for uncoupling analyses of mutations affecting splicing from requirements for transcription and other processing steps. For this purpose, a phage lambda promoter was fused to the Saccharomyces cerevisiae tRNATyr(SUP3a) coding sequence. This fusion ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00240a017

    authors: Shapero MH,Greer CL

    更新日期:1991-07-02 00:00:00

  • Magnesium increases the curvature of duplex DNA that contains dA tracts.

    abstract::Distinct structural features of DNA, such as the curvature of dA tracts, are important in the recognition, packaging, and regulation of DNA. Physiologically relevant concentrations of magnesium have been found to enhance the curvature of dA tract DNAs, as monitored by solution-state NMR, indicating that the structure ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi010853j

    authors: Jerkovic B,Bolton PH

    更新日期:2001-08-07 00:00:00

  • Hematin-hematin self-association states involved in the formation and reactivity of the malaria parasite pigment, hemozoin.

    abstract::The malaria parasite pigment, hemozoin, is a crystal of ferriprotoporphyrin IX (FP-Fe(III)), a product of hemoglobin digestion. Hemozoin formation is essential for FP-Fe(III) detoxification in the parasite; it is the main target of quinoline antimalarials and can modulate immune and inflammation responses. To gain fur...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi100567j

    authors: Klonis N,Dilanian R,Hanssen E,Darmanin C,Streltsov V,Deed S,Quiney H,Tilley L

    更新日期:2010-08-10 00:00:00

  • Structure of pseudobactin A214, a siderophore from a bean-deleterious Pseudomonas.

    abstract::Bean-deleterious Pseudomonas A214 produced the extracellular yellow-green, fluorescent siderophore [microbial iron(III) transport agent] pseudobactin A214 under iron-limiting conditions. Pseudobactin A214 has a molecular formula of C46H64N13O22 and a molecular mass of 1151 g/mol. Pseudobactin A214 contained an N-block...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00367a022

    authors: Buyer JS,Wright JM,Leong J

    更新日期:1986-09-23 00:00:00

  • Signal recognition particle (SRP) and SRP receptor: a new paradigm for multistate regulatory GTPases.

    abstract::The GTP-binding proteins or GTPases comprise a superfamily of proteins that provide molecular switches in numerous cellular processes. The "GTPase switch" paradigm, in which a GTPase acts as a bimodal switch that is turned "on" and "off" by external regulatory factors, has been used to interpret the regulatory mechani...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9006989

    authors: Shan SO,Schmid SL,Zhang X

    更新日期:2009-07-28 00:00:00

  • Characterization of the metal ion binding helix-hairpin-helix motifs in human DNA polymerase beta by X-ray structural analysis.

    abstract::X-ray crystallographic studies have shown that DNA binding by human polymerase beta (pol beta) occurs primarily through two structurally and sequentially homologous helix-hairpin-helix (HhH) motifs, one in the fingers subdomain and the other in the 8-kDa domain [Pelletier, H., Sawaya, M. R., Wolfle, W., Wilson, S. H.,...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi960790i

    authors: Pelletier H,Sawaya MR

    更新日期:1996-10-01 00:00:00

  • Effect of phospholipid oxidation products on transbilayer movement of phospholipids in single lamellar vesicles.

    abstract::Single lamellar phosphatidyl[methyl-2H]choline vesicles were incubated with an excess of unlabeled phosphatidylcholine vesicles or phosphatidylcholine-cholesterol vesicles containing 8 mol % glucuronosyldiglyceride. Incubation of the two vesicle populations was performed in the presence or absence of a purified phosph...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00534a017

    authors: Shaw JM,Thompson TE

    更新日期:1982-03-02 00:00:00

  • Dynamics of backbone conformational heterogeneity in Bacillus subtilis ribonuclease P protein.

    abstract::Interconversion of protein conformations is imperative to function, as evidenced by conformational changes associated with enzyme catalytic cycles, ligand binding and post-translational modifications. In this study, we used 15N NMR relaxation experiments to probe the fast (i.e., ps-ns) and slow (i.e., micros-ms) confo...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi701425n

    authors: Henkels CH,Chang YC,Chamberlin SI,Oas TG

    更新日期:2007-12-25 00:00:00

  • Mutation at active site lysine 212 to arginine uncouples the glycosylase activity from the lyase activity of human endonuclease III.

    abstract::The human endonuclease III (hNTH1) is an important DNA glycosylase with associated abasic lyase activity. We previously demonstrated that the K212Q mutant was totally inactive, while the K212R mutant had reduced DNA glycosylase/lyase activity and could form a covalent complex with the substrate DNA upon reduction. We ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi011053b

    authors: Liu X,Roy R

    更新日期:2001-11-13 00:00:00

  • Conformational Heterogeneity in a Fully Complementary DNA Three-Way Junction with a GC-Rich Branchpoint.

    abstract::DNA three-way junctions (3WJs) are branched structures that serve as important biological intermediates and as components in DNA nanostructures. We recently derived the global structure of a fully complementary 3WJ and found that it contained unpaired bases at the branchpoint, which is consistent with previous observa...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.7b00677

    authors: Toulmin A,Baltierra-Jasso LE,Morten MJ,Sabir T,McGlynn P,Schröder GF,Smith BO,Magennis SW

    更新日期:2017-09-19 00:00:00

  • Structural characterization of azurin from Pseudomonas aeruginosa and some of its methionine-121 mutants.

    abstract::Azurin from Pseudomonas aeruginosa and two mutants where the methionine ligand has been mutated have been studied in order to directly investigate the functional and structural significance of this ligand in the blue copper proteins. Reduction potentials, X-ray absorption fine structure (XAFS), electron paramagnetic r...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00059a013

    authors: Murphy LM,Strange RW,Karlsson BG,Lundberg LG,Pascher T,Reinhammar B,Hasnain SS

    更新日期:1993-03-02 00:00:00

  • Heparin binding domain of antithrombin III: characterization using a synthetic peptide directed polyclonal antibody.

    abstract::Antithrombin III (ATIII) is a plasma-borne serine protease inhibitor that apparently forms covalent complexes with thrombin. The interaction between ATIII and thrombin is enhanced several thousandfold by the glycosaminoglycan, heparin. We have previously proposed that the heparin binding site of ATIII resides within a...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00490a010

    authors: Smith JW,Dey N,Knauer DJ

    更新日期:1990-09-25 00:00:00

  • Functional roles of metal binding domains of the Archaeoglobus fulgidus Cu(+)-ATPase CopA.

    abstract::CopA, a thermophilic membrane ATPase from Archaeoglobus fulgidus, drives the outward movement of Cu(+) or Ag(+) [Mandal et al. (2002) J. Biol. Chem. 277, 7201-7208]. This, as other P(IB)-ATPases, is characterized by a putative metal binding sequence (C(380)PC(382)) in its sixth transmembrane fragment and cytoplasmic m...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi034806y

    authors: Mandal AK,Argüello JM

    更新日期:2003-09-23 00:00:00

  • Transbilayer lipid diffusion promoted by Bax: implications for apoptosis.

    abstract::It is known that the proapoptotic protein Bax facilitates the formation of pores in bilayers, resulting in the release of proteins from the intermitochondrial space. We demonstrate that another consequence of the interaction of Bax with membranes is an increase in the rate of lipid transbilayer diffusion. We use two i...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi035348w

    authors: Epand RF,Martinou JC,Montessuit S,Epand RM

    更新日期:2003-12-16 00:00:00

  • Engineering protein stability and specificity using fluorous amino acids: the importance of packing effects.

    abstract::The incorporation of extensively fluorinated, or fluorous, analogues of hydrophobic amino acids into proteins potentially provides the opportunity to modulate the physicochemical properties of proteins in a predictable manner. On the basis of the properties of small fluorocarbon molecules, extensively fluorinated prot...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi901481k

    authors: Buer BC,de la Salud-Bea R,Al Hashimi HM,Marsh EN

    更新日期:2009-11-17 00:00:00

  • Structural requirements for formyl homooligopeptide chemoattractants.

    abstract::Using solution peptide synthesis, we have made three series of N alpha-formylated homooligopeptides, from the dipeptide to the heptapeptide, derived from L-methionine, L-norleucine, and S-methyl-L-cysteine and related to the chemotactic peptide N alpha-formylmethionylleucylphenylalanine. Compounds were prepared to det...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00299a018

    authors: Toniolo C,Bonora GM,Showell H,Freer RJ,Becker EL

    更新日期:1984-02-14 00:00:00

  • Computational and in vitro analysis of destabilized DNA regions in the interferon gene cluster: potential of predicting functional gene domains.

    abstract::Recent evidence adds support to a traditional concept according to which the eukaryotic nucleus is organized into functional domains by scaffold or matrix attachment regions (S/MARs). These regions have previously been predicted to have a high potential for stress-induced duplex destabilization (SIDD). Here we report ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi026496+

    authors: Goetze S,Gluch A,Benham C,Bode J

    更新日期:2003-01-14 00:00:00

  • Conformation of peptide fragments of proteins in aqueous solution: implications for initiation of protein folding.

    abstract::Applications of sensitive new technologies, in particular, two-dimensional NMR spectroscopy, have allowed detection of folded structures in short peptide fragments of proteins in aqueous solution under conditions where native proteins fold. These structures are in rapid dynamic exchange with unfolded states. These obs...

    journal_title:Biochemistry

    pub_type: 杂志文章,评审

    doi:10.1021/bi00419a001

    authors: Wright PE,Dyson HJ,Lerner RA

    更新日期:1988-09-20 00:00:00

  • Structure-function relationships in sorcin, a member of the penta EF-hand family. Interaction of sorcin fragments with the ryanodine receptor and an Escherichia coli model system.

    abstract::Sorcin, a 21.6 kDa cytosolic EF-hand protein which undergoes a Ca(2+)-induced translocation from cytoplasm to membranes, has been assigned to the newly defined penta EF-hand family. A molecular model of the C-terminal Ca(2+)-binding domain has been generated using as a template the X-ray coordinates of the correspondi...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi991648v

    authors: Zamparelli C,Ilari A,Verzili D,Giangiacomo L,Colotti G,Pascarella S,Chiancone E

    更新日期:2000-02-01 00:00:00

  • Roles of individual domains of annexin I in its vesicle binding and vesicle aggregation: a comprehensive mutagenesis study.

    abstract::To understand the mechanism by which annexin I induces membrane aggregation, a comprehensive mutagenesis of all six Ca2+-binding sites was performed. When the cap residues of type II Ca2+-binding sites were systematically mutated to Ala, a type II site in domain II was shown to be essential for Ca2+-dependent vesicle ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi980479w

    authors: Bitto E,Cho W

    更新日期:1998-07-14 00:00:00

  • Extracellular Juxtamembrane Segment of ADAM17 Interacts with Membranes and Is Essential for Its Shedding Activity.

    abstract::A wide variety of biological processes including differentiation, regeneration, and cancer progression are regulated by shedding of membrane-anchored proteins. One of the major sheddases is A Disintegrin And Metalloprotease-17 (ADAM17) whose extracellular region consists of a pro-, a catalytic, a disintegrin-, and a m...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.5b00497

    authors: Düsterhöft S,Michalek M,Kordowski F,Oldefest M,Sommer A,Röseler J,Reiss K,Grötzinger J,Lorenzen I

    更新日期:2015-09-29 00:00:00

  • The crystal structure of the Escherichia coli maltodextrin phosphorylase-acarbose complex.

    abstract::Acarbose is a naturally occurring pseudo-tetrasaccharide. It has been used in conjunction with other drugs in the treatment of diabetes where it acts as an inhibitor of intestinal glucosidases. To probe the interactions of acarbose with other carbohydrate recognition enzymes, the crystal structure of E. coli maltodext...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9828573

    authors: O'Reilly M,Watson KA,Johnson LN

    更新日期:1999-04-27 00:00:00

  • Amyloid fibrillation of human Apaf-1 CARD.

    abstract::The idea of establishing the amyloid-like fibrillation tendency of pro- and antisurvival proteins of human apoptotic pathways is relevant for delineating the conditions that lead to aberrant differentiation, development, and tissue homeostasis. As the first step in this direction, we report here that the caspase recru...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi900626u

    authors: Rao PN,Reddy KS,Bhuyan AK

    更新日期:2009-08-18 00:00:00

  • 8-Azido-ATP modification of cytochrome c: retardation of its electron-transfer activity to cytochrome c oxidase.

    abstract::Horse heart cytochrome c has been modified by 8-azido-ATP and the electron-transfer activity of the modified cytochrome c's to bovine heart cytochrome c oxidase (CcO) under physiological ionic strengths has been studied by the laser flash photolysis technique with 5-deazariboflavin and EDTA as the electron donor. The ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00008a035

    authors: Lin J,Wu S,Lau WT,Chan SI

    更新日期:1995-02-28 00:00:00

  • Two homologues encoding human UDP-glucose:glycoprotein glucosyltransferase differ in mRNA expression and enzymatic activity.

    abstract::UDP-glucose:glycoprotein glucosyltransferase (UGT) is a soluble protein of the endoplasmic reticulum (ER) that operates as a gatekeeper for quality control by preventing transport of improperly folded glycoproteins out of the ER. We report the isolation of two cDNAs encoding human UDP-glucose:glycoprotein glucosyltran...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9916473

    authors: Arnold SM,Fessler LI,Fessler JH,Kaufman RJ

    更新日期:2000-03-07 00:00:00

  • The flavoprotein component of the Escherichia coli sulfite reductase: expression, purification, and spectral and catalytic properties of a monomeric form containing both the flavin adenine dinucleotide and the flavin mononucleotide cofactors.

    abstract::The flavoprotein component (SiR-FP) of the sulfite reductase from Escherichia coli is an octamer containing one FAD and one FMN per polypeptide chain. SiR-FP60, a SiR-FP fragment starting with alanine-52, was overexpressed in E. coli and purified as a monomer. The N-terminal part of the native protein contains thus al...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9728699

    authors: Zeghouf M,Fontecave M,Macherel D,Covès J

    更新日期:1998-04-28 00:00:00

  • NMR structural studies of a 15-mer DNA sequence from a ras protooncogene, modified at the first base of codon 61 with the carcinogen 4-aminobiphenyl.

    abstract::Proton NMR studies were conducted on the complementary 15-mer duplex d(5'-TACTCTTCTTGACCT).(5'-AGGTCAAGAAGAGTA) (designated as unmodified 15-mer duplex) spanning a portion of the mouse c-Ha-ras protooncogene centered around codon 61. Identical studies were carried out on the same sequence, after specific modification ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00155a011

    authors: Cho BP,Beland FA,Marques MM

    更新日期:1992-10-13 00:00:00