当前位置： SCI文献检索 > BIOCHEMISTRY期刊下所有文献 > Spectroscopic evidence that osmolytes used in crystallization buffers inhibit a conformation change in a membrane protein.

Spectroscopic evidence that osmolytes used in crystallization buffers inhibit a conformation change in a membrane protein.

Abstract：

:BtuB is a bacterial outer-membrane protein that transports vitamin B(12). Spectroscopic studies using site-directed spin labeling (SDSL) indicate that the N-terminus of BtuB undergoes a dramatic structural change from a docked (folded) to an undocked (unfolded) configuration upon substrate binding. However, this dramatic conformational change is not observed in the crystal structures of BtuB. Here, we make an attempt to resolve this discrepancy and find that the effects of solutes can explain the discrepancy between the results obtained using these two methods. Specifically, if SDSL is performed with the buffers used for the crystallization of BtuB, the substrate-induced order-disorder transition of the N-terminal Ton box observed in intact membranes is blocked. Moreover, poly(ethylene glycol) 3350, which is a component of the crystallization and soaking buffers, is shown to inhibit this structural transition. It is likely that the crystal structure of BtuB in its holo form represents an osmotically trapped conformation. Conformational changes involving relatively modest energy differences and significant hydration changes may be sensitive to solutes used during crystallization, and this example demonstrates the value of combining multiple structural methods in the examination of protein structure and function.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Fanucci GE,Lee JY,Cafiso DS

doi

10.1021/bi035439t

subject

Has Abstract

pub_date

2003-11-18 00:00:00

pages

13106-12

issue

45

eissn

0006-2960

issn

1520-4995

journal_volume

42

pub_type

杂志文章

相关文献

BIOCHEMISTRY文献大全
  • Zn(2+)-dependent misfolding of the p53 DNA binding domain.

    abstract：:The DNA binding domain (DBD) of p53 folds by a complex mechanism that involves parallel pathways and multiple intermediates, both on- and off-pathway. This heterogeneity renders DBD particularly susceptible to misfolding and aggregation. The origins of parallel folding mechanisms are not well understood. DBD folding h...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi062106y

    authors： Butler JS,Loh SN

    更新日期：2007-03-13 00:00:00

  • HlyC, the internal protein acyltransferase that activates hemolysin toxin: the role of conserved tyrosine and arginine residues in enzymatic activity as probed by chemical modification and site-directed mutagenesis.

    abstract：:Internal fatty acylation of proteins is a recognized means of modifying biological behavior. Escherichia coli hemolysin A (HlyA), a toxic protein, is transcribed as a nontoxic protein and made toxic by internal acylation of two lysine residue epsilon-amino groups; HlyC catalyzes the acyl transfer from acyl-acyl carrie...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi990138y

    authors： Trent MS,Worsham LM,Ernst-Fonberg ML

    更新日期：1999-07-06 00:00:00

  • Site-directed mutagenesis of kappa-bungarotoxin: implications for neuronal receptor specificity.

    abstract：:Postsynaptic polypeptide neurotoxins isolated from the venoms of elapid and hydrophid snakes exhibit the ability to bind selectively to and inhibit different types of receptors that function in nerve signal transmission. On the basis of their amino acid sequences and three-dimensional structures, these neurotoxins are...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00179a011

    authors： Fiordalisi JJ,al-Rabiee R,Chiappinelli VA,Grant GA

    更新日期：1994-04-05 00:00:00

  • Probing Structural Dynamics and Topology of the KCNE1 Membrane Protein in Lipid Bilayers via Site-Directed Spin Labeling and Electron Paramagnetic Resonance Spectroscopy.

    abstract：:KCNE1 is a single transmembrane protein that modulates the function of voltage-gated potassium channels, including KCNQ1. Hereditary mutations in the genes encoding either protein can result in diseases such as congenital deafness, long QT syndrome, ventricular tachyarrhythmia, syncope, and sudden cardiac death. Despi...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.5b00505

    authors： Sahu ID,Craig AF,Dunagan MM,Troxel KR,Zhang R,Meiberg AG,Harmon CN,McCarrick RM,Kroncke BM,Sanders CR,Lorigan GA

    更新日期：2015-10-20 00:00:00

  • Mutational analysis of the "regulatory module" of hormone-sensitive lipase.

    abstract：:Hormone-sensitive lipase (HSL) is a rate-limiting enzyme in lipolysis that displays broad substrate specificity. HSL function is regulated by reversible phosphorylation that occurs within a 150 aa "regulatory module" of the protein. The current studies used mutational analysis to dissect the contribution of the "regul...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi049206t

    authors： Wang J,Shen WJ,Patel S,Harada K,Kraemer FB

    更新日期：2005-02-15 00:00:00

  • Complete assignment of disulfide bonds in bovine dopamine beta-hydroxylase.

    abstract：:Peptide mapping, chemical sequencing, microbore HPLC/electrospray ionization mass spectrometry (LC/ESI/MS), and matrix-assisted laser desorption mass spectrometry (MALDI/MS) were used to identify the sites of intra- and intermolecular disulfide linkages in bovine dopamine beta-hydroxylase. The enzyme contains 14 cyste...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00204a019

    authors： Robertson JG,Adams GW,Medzihradszky KF,Burlingame AL,Villafranca JJ

    更新日期：1994-09-27 00:00:00

  • Structure of the conserved HAMP domain in an intact, membrane-bound chemoreceptor: a disulfide mapping study.

    abstract：:The HAMP domain is a conserved motif widely distributed in prokaryotic and lower eukaryotic organisms, where it is often found in transmembrane receptors that regulate two-component signaling pathways. The motif links receptor input and output modules and is essential to receptor structure and signal transduction. Rec...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi701832b

    authors： Swain KE,Falke JJ

    更新日期：2007-12-04 00:00:00

  • Identification of a novel noncatalytic bicarbonate binding site in eubacterial beta-carbonic anhydrase.

    abstract：:The structures of beta class carbonic anhydrases (beta-CAs) determined so far fall into two distinct subclasses based on the observed coordination of the catalytic zinc (Zn2+) ion. The subclass of beta-CAs that coordinate Zn2+ tetrahedrally with four protein-derived ligands is represented by the structures of ortholog...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi052272q

    authors： Cronk JD,Rowlett RS,Zhang KY,Tu C,Endrizzi JA,Lee J,Gareiss PC,Preiss JR

    更新日期：2006-04-11 00:00:00

  • Characterization of a protein-generated O₂ binding pocket in PqqC, a cofactorless oxidase catalyzing the final step in PQQ production.

    abstract：:PQQ is an exogenous, tricyclic, quino-cofactor for a number of bacterial dehydrogenases. The final step of PQQ formation is catalyzed by PqqC, a cofactorless oxidase. This study focuses on the activation of molecular oxygen in an enzyme active site without metal or cofactor and has identified a specific oxygen binding...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi1015474

    authors： RoseFigura JM,Puehringer S,Schwarzenbacher R,Toyama H,Klinman JP

    更新日期：2011-03-08 00:00:00

  • Methylated pyrimidines stabilize an alternating conformation of poly(dA-dU).poly(dA-dU).

    abstract：:We have investigated the effect of increasing percentages of methylated pyrimidines on the structure of poly(dA-dU).poly(dA-dU). This was done by synthesizing analogous polynucleotides that contained deoxythymidine residues as well as deoxyuridine residues and observing their 31P NMR spectra in increasing amounts of C...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00341a033

    authors： Wu HY,Behe MJ

    更新日期：1985-09-24 00:00:00

  • The purple to blue transition of bacteriorhodopsin is accompanied by a loss of the hexagonal lattice and a conformational change.

    abstract：:X-ray diffraction measurements show that in contrast to the purple membrane, the bacteriorhodopsin molecules are not organized in a hexagonal lattice in the deionized blue membrane. Addition of Ca2+ restores both the purple color and the normal (63 A) hexagonal protein lattice. In the blue state, the circular dichrois...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00449a031

    authors： Heyn MP,Dudda C,Otto H,Seiff F,Wallat I

    更新日期：1989-11-14 00:00:00

  • A method for assessing the stability of a membrane protein.

    abstract：:The integral membrane protein diacylglycerol kinase (DGK) from Escherichia coli has been reversibly unfolded in a protein/detergent/mixed micelle system by varying the molar ratio of n-decyl beta-D-maltoside (DM) and sodium dodecyl sulfate (SDS). Unfolding was monitored by circular dichroism (CD) and ultraviolet (UV) ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi963095j

    authors： Lau FW,Bowie JU

    更新日期：1997-05-13 00:00:00

  • The ionic track in the F1-ATPase from the thermophilic Bacillus PS3.

    abstract：:Only beta-beta cross-links form when the alpha(3)(betaE(395)C)(3)gammaK(36)C (MF(1) residue numbers) double mutant subcomplex of TF(1), the F(1)-ATPase from the thermophilic Bacillus PS3, is slowly inactivated with CuCl(2) in the presence or absence of MgATP. The same slow rate of inactivation and extent of beta-beta ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi036058i

    authors： Bandyopadhyay S,Allison WS

    更新日期：2004-03-09 00:00:00

  • (15)N NMR relaxation data reveal significant chemical exchange broadening in the alpha-domain of human alpha-lactalbumin.

    abstract：:Human alpha-lactalbumin (alpha-LA), a 123-residue calcium-binding protein, has been studied using (15)N NMR relaxation methods in order to characterize backbone dynamics of the native state at the level of individual residues. Relaxation data were collected at three magnetic field strengths and analyzed using the mode...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi900023m

    authors： Bruylants G,Redfield C

    更新日期：2009-05-19 00:00:00

  • Chemistry Is Dead. Long Live Chemistry!

    abstract：:Chemistry, once king of fluorescence microscopy, was usurped by the field of fluorescent proteins. The increased demands of modern microscopy techniques on the "photon budget" require better and brighter fluorophores, causing a renewed interest in synthetic dyes. Here, we review the recent advances in biochemistry, pr...

    journal_title：Biochemistry

    pub_type： 杂志文章,评审

    doi：10.1021/acs.biochem.7b00529

    authors： Lavis LD

    更新日期：2017-10-03 00:00:00

  • Structure-sensitive RNA footprinting of yeast nuclear ribonuclease P.

    abstract：:Several enzymatic and chemical reagents were used to probe the secondary structure of Saccharomyces cerevisiae nuclear RNase P RNA in the presence and absence of its protein components. Double-stranded regions were detected with RNase V1 and single-stranded regions with RNase ONE (Escherichia coli RNase I). Nucleotide...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00173a022

    authors： Tranguch AJ,Kindelberger DW,Rohlman CE,Lee JY,Engelke DR

    更新日期：1994-02-22 00:00:00

  • Formation and rejoining of deoxyribonucleic acid double-strand breaks induced in isolated cell nuclei by antineoplastic intercalating agents.

    abstract：:The biochemical characteristics of the formation and disappearance of intercalator-induced DNA double-strand breaks (DSB) were studied in nuclei from mouse leukemia L1210 cells by using filter elution methodology [Bradley, M. O., & Kohn, K.W. (1979) Nucleic Acids Res. 7, 793-804]. The three intercalators used were 4'-...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00309a013

    authors： Pommier Y,Schwartz RE,Kohn KW,Zwelling LA

    更新日期：1984-07-03 00:00:00

  • 31P NMR visibility and characterization of rat liver mitochondrial matrix adenine nucleotides.

    abstract：:Compartmentation and NMR visibility of mitochondrial adenine nucleotides were quantitated in isolated rat liver mitochondria respiring on succinate and glutamate in vitro at 8 and 25 degrees C. Intra- and extramitochondrial nucleotides were discriminated by adding the chelator trans-1,2-diaminocyclohexane-N,N,N',N'-te...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00436a030

    authors： Hutson SM,Berkich D,Williams GD,LaNoue KF,Briggs RW

    更新日期：1989-05-16 00:00:00

  • Mechanistic and metabolic inferences from the binding of substrate analogues and products to arginase.

    abstract：:Arginase is a binuclear Mn(2+) metalloenzyme that catalyzes the hydrolysis of L-arginine to L-ornithine and urea. X-ray crystal structures of arginase complexed to substrate analogues N(omega)-hydroxy-L-arginine and N(omega)-hydroxy-nor-L-arginine, as well as the products L-ornithine and urea, complete a set of struct...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi002318+

    authors： Cox JD,Cama E,Colleluori DM,Pethe S,Boucher JL,Mansuy D,Ash DE,Christianson DW

    更新日期：2001-03-06 00:00:00

  • The crystal structure of eosinophil cationic protein in complex with 2',5'-ADP at 2.0 A resolution reveals the details of the ribonucleolytic active site.

    abstract：:Eosinophil cationic protein (ECP) is a component of the eosinophil granule matrix. It shows marked toxicity against helminth parasites, bacteria single-stranded RNA viruses, and host epithelial cells. Secretion of human ECP is related to eosinophil-associated allergic, asthmatic, and inflammatory diseases. ECP belongs...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi0264521

    authors： Mohan CG,Boix E,Evans HR,Nikolovski Z,Nogués MV,Cuchillo CM,Acharya KR

    更新日期：2002-10-08 00:00:00

  • Proposed structural mechanism of Escherichia coli cAMP receptor protein cAMP-dependent proteolytic cleavage protection and selective and nonselective DNA binding.

    abstract：:The Escherichia coli cAMP receptor protein (CRP) displays biphasic characteristics in protease and beta-galactosidase induction assays at increasing cAMP concentrations in response to ligand binding at the secondary binding site located between the primary binding site and the DNA binding domain. Two mutants were crea...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi0479609

    authors： Scott SP,Jarjous S

    更新日期：2005-06-21 00:00:00

  • Cyanide binding at the non-heme Fe2+ of the iron-quinone complex of photosystem II: at high concentrations, cyanide converts the Fe2+ from high (S = 2) to low (S = 0) spin.

    abstract：:The primary electron acceptor complex of photosystem II, QAFe2+, can bind a number of small molecules at the iron site, including cyanide [Koulougliotis, D., Kostopoulos, T., Petrouleas, V., & Diner, B. A. (1993) Biochim. Biophys. Acta 1141, 275-282)]. In the presence of NaCN (30-300 mM) at pH 6.5, the reduced state, ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00199a014

    authors： Sanakis Y,Petrouleas V,Diner BA

    更新日期：1994-08-23 00:00:00

  • Hydrolysis of proteins using dipeptidyl aminopeptidases: analysis of the N-terminal portion of spinach plastocyanin.

    abstract：:The exopeptidases dipeptidyl aminopeptidases I and IV were used to hydrolyze the N-terminal portion of spinach plastocyanin to dipeptides. The enzymes were used individually as well as in a mixture and the dipeptides were analyzed by combined gas chromatography-mass spectrometry. Data are presented for native plastocy...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00596a009

    authors： Seifert WE Jr,Caprioli RM

    更新日期：1978-02-07 00:00:00

  • An endogenous sulfated inhibitor of neuronal inositol trisphosphate receptors.

    abstract：:In cerebellum, inositol trisphosphate- (InsP(3)-) gated Ca channels play a key role in learning, though they exhibit a low sensitivity to InsP(3) compared to peripheral tissues. In the present study, the cerebellar InsP(3) receptor is shown to be associated with a novel inhibitor of InsP(3) binding. (3)H-InsP(3) bindi...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi992175v

    authors： Watras J,Orlando R,Moraru II

    更新日期：2000-03-28 00:00:00

  • Acyclic guanosine analogs inhibit DNA polymerases alpha, delta, and epsilon with very different potencies and have unique mechanisms of action.

    abstract：:Acyclovir triphosphate, ganciclovir triphosphate and penciclovir triphosphate inhibited DNA polymerases alpha, delta, and epsilon. Each triphosphate preferentially inhibited pol delta, although ganciclovir triphosphate was the most impressive of the three; the Ki for inhibition of pol delta was 2 microM (competitive w...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00008a014

    authors： Ilsley DD,Lee SH,Miller WH,Kuchta RD

    更新日期：1995-02-28 00:00:00

  • Resonance Raman characterization of the heme cofactor in cystathionine beta-synthase. Identification of the Fe-S(Cys) vibration in the six-coordinate low-spin heme.

    abstract：:Human cystathionine beta-synthase (CBS) is an essential enzyme for the removal of the toxic metabolite homocysteine. Heme and pyridoxal phosphate (PLP) cofactors are necessary to catalyze the condensation of homocysteine and serine to generate cystathionine. While the role for the PLP cofactor is thought to be similar...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi0010874

    authors： Green EL,Taoka S,Banerjee R,Loehr TM

    更新日期：2001-01-16 00:00:00

  • Identification of 9-cis,13-cis-retinoic acid as a major circulating retinoid in plasma.

    abstract：:In the course of measuring the concentrations of retinoic acids (RA) in bovine plasma, a major peak was observed which comigrated with 9-cis-RA on normal-phase high-performance liquid chromatography. Rechromatography of this retinoic on reverse-phase high-performance liquid chromatography showed that it was distinct f...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00004a013

    authors： Horst RL,Reinhardt TA,Goff JP,Nonnecke BJ,Gambhir VK,Fiorella PD,Napoli JL

    更新日期：1995-01-31 00:00:00

  • Role of the conserved phenylalanine 181 of NADPH-cytochrome P450 oxidoreductase in FMN binding and catalytic activity.

    abstract：:NADPH-cytochrome P450 oxidoreductase (P450 reductase, EC 1.6.2.4) is an essential component of the P450 monooxygenase complex and binds FMN, FAD, and NADPH cofactors. Residues Tyr140 and Tyr178 are known to be involved in FMN binding. A third aromatic side chain, Phe181, is also located in the proximity of the FMN rin...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi011147l

    authors： Paine MJ,Ayivor S,Munro A,Tsan P,Lian LY,Roberts GC,Wolf CR

    更新日期：2001-11-13 00:00:00

  • Glutathione Directly Intercepts DNA Radicals To Inhibit Oxidative DNA-Protein Cross-Linking Induced by the One-Electron Oxidation of Guanine.

    abstract：:Oxidative DNA damage can lead to cancer, and as enzymatic DNA repair systems become compromised during the aging process, the role of exogenous antioxidants becomes more critical. Here, we examined whether such non-enzymatic DNA repair can be effected by the common cellular antioxidant glutathione, investigating both ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.9b00697

    authors： Safaeipour M,Jauregui J,Castillo S,Bekarian M,Esparza D,Sanchez M,Stemp EDA

    更新日期：2019-11-19 00:00:00

  • Influence of histidine tag attachment on picosecond protein dynamics.

    abstract：:Polyhistidine affinity tags are routinely employed as a convenient means of purifying recombinantly expressed proteins. A tacit assumption is commonly made that His tags have little influence on protein structure and function. Attachment of a His tag to the N-terminus of the robust globular protein myoglobin leads to ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi2003923

    authors： Thielges MC,Chung JK,Axup JY,Fayer MD

    更新日期：2011-06-28 00:00:00