Spin-label and fluorescence labeling studies of the thioester bonds in human alpha 2-macroglobulin.


:Upon cleavage of the reactive thioester bonds (Cys-949-Glx-952) of tetrameric human alpha 2-macroglobulin (alpha 2M) by methylamine, one sulfhydryl group per alpha 2M subunit is exposed. These identical sulfhydryl group sites were labeled with the thiol-specific nitroxide spin-labels (1-oxy-2,2,5,5-tetramethyl-3-pyrrolin-3-yl)methyl methanethiosulfonate and (1-oxy-2,2,6,6-tetramethyl-4-piperidinyl)methyl methanethiosulfonate, a homologous series of maleimide spin-labels, and the thiol-specific fluorescent probe 2-[(4-maleimidophenyl)amino]naphthalene-6-sulfonic acid sodium salt (MANS). The ESR and fluorescence results showed that these sulfhydryl group sites were at the base of a narrow crevice that is greater than or equal to 8 A deep. Although the bound MANS fluorophore was slightly blue shifted with an enhanced quantum yield vs the free label in water, the environment of the sulfhydryl site appeared to be of a polar nature when compared with the emission maxima in several solvents of varying polarity. The Glx residue participating in the thioester linkage in the intact protein was labeled with 4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl. The distance between the Glx and Cys moieties was estimated at greater than or equal to 10-25 A from double spin-labeling experiments.






Zhao BL,Musci G,Sugawara Y,Berliner LJ




Has Abstract


1988-07-12 00:00:00












  • Nuclear magnetic resonance and neutron diffraction studies of the complex of ribonuclease A with uridine vanadate, a transition-state analogue.

    abstract::The complex of ribonuclease A (RNase A) with uridine vanadate (U-V), a transition-state analogue, has been studied with 51V and proton NMR spectroscopy in solution and by neutron diffraction in the crystalline state. Upon the addition of aliquots of U-V at pH 6.6, the C epsilon-H resonances of the two active-site hist...


    pub_type: 杂志文章


    authors: Borah B,Chen CW,Egan W,Miller M,Wlodawer A,Cohen JS

    更新日期:1985-04-09 00:00:00

  • The activating transcription factor region within the E2A promoter exists in a novel conformation.

    abstract::The ATF (activating transcription factor) binding site within the E2A promoter region of adenovirus is shown to exist in a novel conformation in vitro via nuclear magnetic resonance methods. This novel conformation may be important to the protein DNA recognition process. This conformation has characteristics of both A...


    pub_type: 杂志文章


    authors: Borden KL

    更新日期:1993-07-06 00:00:00

  • Cofactor Complexes of DesD, a Model Enzyme in the Virulence-related NIS Synthetase Family.

    abstract::The understudied nonribosomal-peptide-synthetase-independent siderophore (NIS) synthetase family has been increasingly associated with virulence in bacterial species due to its key role in the synthesis of hydroxamate and carboxylate "stealth" siderophores. We have identified a model family member, DesD, from Streptom...


    pub_type: 杂志文章


    authors: Hoffmann KM,Goncuian ES,Karimi KL,Amendola CR,Mojab Y,Wood KM,Prussia GA,Nix J,Yamamoto M,Lathan K,Orion IW

    更新日期:2020-09-22 00:00:00

  • The high molecular weight nerve growth factor complex from Mastomys natalensis differs from the murine nerve growth factor complex.

    abstract::Both the male and the female of Mastomys natalensis, an African rat, have high levels of nerve growth factor (NGF) in their submaxillary glands. Mastomys high molecular weight (HMW) NGF was purified by gel filtration and ion-exchange chromatography and was compared with HMW NGF from the male mouse submaxillary gland. ...


    pub_type: 杂志文章


    authors: Darling TL,Fahnestock M

    更新日期:1988-09-06 00:00:00

  • Progressive development of a thrombin inhibitor binding site.

    abstract::The studies reported here were undertaken to determine whether the thrombin precursors prothrombin, prethrombin 1, prethrombin 2, and Meizo thrombin interact with the fluorescent, reversible thrombin inhibitor dansylarginine N,N-(3-ethyl-1,5-pentanediyl)amide (DAPA) [Nesheim, M. E., Prendergast, F. G., & Mann, K. G. (...


    pub_type: 杂志文章


    authors: Hibbard LS,Nesheim ME,Mann KG

    更新日期:1982-05-11 00:00:00

  • Analysis of folding and unfolding reactions of cytochrome b5.

    abstract::The guanidine hydrochloride- (GuHCl-) induced unfolding and refolding of a recombinant domain of bovine microsomal cytochrome b(5) containing the first 104 amino acid residues has been characterized by both transient and equilibrium spectrophotometric methods. The soluble domain is reversibly unfolded and the equilibr...


    pub_type: 杂志文章


    authors: Manyusa S,Mortuza G,Whitford D

    更新日期:1999-10-26 00:00:00

  • Kinetics of long-chain (sphingoid) base biosynthesis in intact LM cells: effects of varying the extracellular concentrations of serine and fatty acid precursors of this pathway.

    abstract::Serine palmitoyltransferase (EC catalyzes the condensation of L-serine and palmitoyl-CoA to yield 3-ketosphinganine in the first unique reaction of long-chain (sphingoid) base biosynthesis. The kinetic effects of changing the extracellular concentrations of the precursors for this pathway were studied with L...


    pub_type: 杂志文章


    authors: Merrill AH Jr,Wang E,Mullins RE

    更新日期:1988-01-12 00:00:00

  • Uniform 13C isotope labeling of proteins with sodium acetate for NMR studies: application to human carbonic anhydrase II.

    abstract::Uniform double labeling of proteins for NMR studies can be prohibitively expensive, even with an efficient expression and purification scheme, due largely to the high cost of [13C6, 99%]glucose. We demonstrate here that uniformly (greater than 95%) 13C and 15N double-labeled proteins can be prepared for NMR structure/...


    pub_type: 杂志文章


    authors: Venters RA,Calderone TL,Spicer LD,Fierke CA

    更新日期:1991-05-07 00:00:00

  • Hydrophobic core of the steroidogenic acute regulatory protein for cholesterol transport.

    abstract::The steroidogenic acute regulatory protein (StAR), the first family member of START (StAR-related lipid transport) proteins, plays an essential role by facilitating the movement of cholesterol from the outer to inner mitochondrial membrane. Wild-type and mutant StAR binds cholesterol with similar intensity, but only w...


    pub_type: 杂志文章


    authors: Bose HS,Whittal RM,Bose M,Debnath D

    更新日期:2009-02-17 00:00:00

  • Identification by proton nuclear magnetic resonance of the histidines in cytochrome b5 modified by diethyl pyrocarbonate.

    abstract::Diethyl pyrocarbonate (DEP) is an electrophilic reagent that is used to modify reversibly the histidine residues of proteins. Unfortunately, the lability of the acylated histidine adduct usually does not permit the isolation and identification of the modified histidine. By use of 500-MHz proton NMR spectroscopy, it ha...


    pub_type: 杂志文章


    authors: Altman J,Lipka JJ,Kuntz I,Waskell L

    更新日期:1989-09-19 00:00:00

  • Critical micelle concentrations of gangliosides.

    abstract::The micellar properties of mixed, bovine gangliosides and purified galactosyl-N-acetylacetylgalactosaminyl (N-acetylneuraminyl) galactosylglucosylceramide were studied by gel filtration, equilibrium dialysis, and band and boundary centrifugation in sucrose gradients. The dissociation of micelles is very slow (days) in...


    pub_type: 杂志文章


    authors: Formisano S,Johnson ML,Lee G,Aloj SM,Edelhoch H

    更新日期:1979-03-20 00:00:00

  • Intersubunit communication in tryptophan synthase by carbon-13 and fluorine-19 REDOR NMR.

    abstract::The beta subunits of the 143-kDa alpha2beta2 tetrameric enzyme tryptophan synthase have been labeled by L-[ring-4-19F]phenylalanine and L-[phenol-4-13C]tyrosine in an effort to monitor the positions of these residues on ligand binding. Of the 13 phenylalanine and 11 tyrosine residues in the beta subunit, only three pa...


    pub_type: 杂志文章


    authors: McDowell LM,Lee M,McKay RA,Anderson KS,Schaefer J

    更新日期:1996-03-12 00:00:00

  • Approaches to predicting effects of single amino acid substitutions on the function of a protein.

    abstract::The relative activities of 313 mutants of the gene V protein of bacteriophage f1, assayed in vivo, have been used to evaluate two approaches to predicting the effects of single amino acid substitutions on the function of a protein. First, we tested methods that only depend on the properties of the wild-type and substi...


    pub_type: 杂志文章


    authors: Zabin HB,Horvath MP,Terwilliger TC

    更新日期:1991-06-25 00:00:00

  • Abortive initiation and long ribonucleic acid synthesis.

    abstract::In vitro transcription assays have been used to study the rate of ribonucleic acid (RNA) synthesis from the Escherichia coli lactose promotor mutant lacL8UV5 contained on a 203-bp (base pair) restriction fragment. The half-life of long (63-base) RNA production from heparin-resistant RNA polymerase-promotor complexes w...


    pub_type: 杂志文章


    authors: Munson LM,Reznikoff WS

    更新日期:1981-04-14 00:00:00

  • CHOBIMALT: a cholesterol-based detergent.

    abstract::Cholesterol and its hemisuccinate and sulfate derivatives are widely used in studies of purified membrane proteins but are difficult to solubilize in aqueous solution, even in the presence of detergent micelles. Other cholesterol derivatives do not form conventional micelles and lead to viscous solutions. To address t...


    pub_type: 杂志文章


    authors: Howell SC,Mittal R,Huang L,Travis B,Breyer RM,Sanders CR

    更新日期:2010-11-09 00:00:00

  • 4-Azido-2-nitrophenyl phosphate, a new photoaffinity derivative of inorganic phosphate. Study of its interaction with the inorganic phosphate binding site of beef heart mitochondrial adenosine triphosphatase.

    abstract::4-Azido-2-nitrophenyl phosphate (ANPP) was synthesized and characterized. ANPP, unlabeled or labeled by 32P, was used as a photoreactive analogue of Pi to study the Pi binding site(s) in isolated F1-ATPase and inside-out particles from beef heart mitochondria. In the dark, the phosphate bond of ANPP was cleaved by alk...


    pub_type: 杂志文章


    authors: Lauquin G,Pougeois R,Vignais PV

    更新日期:1980-09-30 00:00:00

  • Structural origins of the functional divergence of human insulin-like growth factor-I and insulin.

    abstract::Human insulin-like growth factors I and II (hIGF-I, hIGF-II) are potent stimulators of cell and growth processes. They display high sequence similarity to both the A and B chains of insulin but contain an additional connecting C-domain, which reflects their secretion without specific packaging or precursor conversion....


    pub_type: 杂志文章


    authors: Brzozowski AM,Dodson EJ,Dodson GG,Murshudov GN,Verma C,Turkenburg JP,de Bree FM,Dauter Z

    更新日期:2002-07-30 00:00:00

  • Interpretation of the doublet at 850 and 830 cm-1 in the Raman spectra of tyrosyl residues in proteins and certain model compounds.

    abstract::The doublet at 850 and 830 cm-1 in the Raman spectra of proteins containing tyrosyl residues has been examined as to its origin and the relation of its components to the environment of the phenyl ring, the state of the phenolic hydroxyl group, and the conformation of the amino acid backbone. Raman spectral studies on ...


    pub_type: 杂志文章


    authors: Siamwiza MN,Lord RC,Chen MC,Takamatsu T,Harada I,Matsuura H,Shimanouchi T

    更新日期:1975-11-04 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...


    pub_type: 杂志文章


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

    更新日期:2003-12-16 00:00:00

  • Surface plasmon resonance binding kinetics of Alzheimer's disease amyloid beta peptide-capturing and plaque-binding monoclonal antibodies.

    abstract::Several different monoclonal antibodies (mAbs) have been actively developed in the field of Alzheimer's disease (AD) for basic science and clinical applications; however, the binding kinetics of many of the mAbs with the beta-amyloid peptides (Abeta) are poorly understood. A panel of mAbs with different Abeta recognit...


    pub_type: 杂志文章


    authors: Ramakrishnan M,Kandimalla KK,Wengenack TM,Howell KG,Poduslo JF

    更新日期:2009-11-03 00:00:00

  • Three-dimensional structure of murine anti-p-azophenylarsonate Fab 36-71. 2. Structural basis of hapten binding and idiotypy.

    abstract::Comparison between the structures and solvent-accessible surfaces of the antigen-binding fragments of two murine anti-p-azophenylarsonate monoclonal antibodies, one bearing a major cross-reactive idiotype of A/J strain mice (36-71) and one lacking the idiotype (R19.9; Lascombe et al., 1989), highlight the structural b...


    pub_type: 杂志文章


    authors: Strong RK,Petsko GA,Sharon J,Margolies MN

    更新日期:1991-04-16 00:00:00

  • Purification and characterization of a transmembrane domain-deleted form of lecithin retinol acyltransferase.

    abstract::Lecithin retinol acyltransferase (LRAT) catalyzes the esterification of all-trans-retinol into all-trans-retinyl ester, an essential reaction in the vertebrate visual cycle. Since all-trans-retinyl esters are the substrates for the isomerization reaction that generates 11-cis-retinoids, this esterification reaction is...


    pub_type: 杂志文章


    authors: Bok D,Ruiz A,Yaron O,Jahng WJ,Ray A,Xue L,Rando RR

    更新日期:2003-05-27 00:00:00

  • The BARD1 C-terminal domain structure and interactions with polyadenylation factor CstF-50.

    abstract::The BARD1 N-terminal RING domain binds BRCA1 while the BARD1 C-terminal ankyrin and tandem BRCT repeat domains bind CstF-50 to modulate mRNA processing and RNAP II stability in response to DNA damage. Here we characterize the BARD1 structural biochemistry responsible for CstF-50 binding. The crystal structure of the B...


    pub_type: 杂志文章


    authors: Edwards RA,Lee MS,Tsutakawa SE,Williams RS,Nazeer I,Kleiman FE,Tainer JA,Glover JN

    更新日期:2008-11-04 00:00:00

  • Investigating the in Vitro Thermal Stability and Conformational Flexibility of Estrogen Receptors as Potential Key Factors of Their in Vivo Activity.

    abstract::Among hormone-inducible transcription factors, estrogen receptors (ERs) play important roles in tissue growth and differentiation, via either direct or indirect binding, in the nucleus, to specific DNA targets called estrogen responsive elements (EREs), or through nongenomic pathways. In humans, two estrogen receptor ...


    pub_type: 杂志文章


    authors: Le Grand A,André-Leroux G,Marteil G,Duval H,Sire O,Le Tilly V

    更新日期:2015-06-30 00:00:00

  • Effect of sequence on the conformation of DNA holliday junctions.

    abstract::Structures of the DNA sequences d(CCGGCGCCGG) and d(CCAGTACbr(5)UGG) are presented here as four-way Holliday junctions in their compact stacked-X forms, with antiparallel alignment of the DNA strands. Thus, the ACC-trinucleotide motif, previously identified as important for stabilizing the junction, is now extended to...


    pub_type: 杂志文章


    authors: Hays FA,Vargason JM,Ho PS

    更新日期:2003-08-19 00:00:00

  • The role of phenylalanine 31 in maintaining the conformational stability of ribonuclease P2 from Sulfolobus solfataricus under extreme conditions of temperature and pressure.

    abstract::Ribonuclease P2 from the thermophilic archaebacterium Sulfolobus solfataricus is a small protein (7 kDa) with a known three-dimensional structure. Inspection of the structure and molecular dynamics simulation reveal that three aromatic residues (Phe5, Phe31, and Tyr33) from the hydrophobic core have a strong van der W...


    pub_type: 杂志文章


    authors: Mombelli E,Afshar M,Fusi P,Mariani M,Tortora P,Connelly JP,Lange R

    更新日期:1997-07-22 00:00:00

  • Solid-state nuclear magnetic resonance evidence for parallel and antiparallel strand arrangements in the membrane-associated HIV-1 fusion peptide.

    abstract::The HIV-1 fusion peptide serves as a useful model system for understanding viral/target cell fusion, at least to the lipid-mixing stage. Previous solid-state NMR studies have shown that the membrane-bound HIV-1 fusion peptide adopts an extended conformation in a lipid mixture close to that of host cells of the virus. ...


    pub_type: 杂志文章


    authors: Yang J,Weliky DP

    更新日期:2003-10-14 00:00:00

  • Characterization of microtubule-associated protein MAP1B: phosphorylation state, light chains, and binding to microtubules.

    abstract::We have recently described a procedure for the purification of microtubule associated protein 1B (MAP1B) from calf brain [Pedrotti, B., & Islam K. (1995) Cell Motil. Cytoskeleton 30, 301-309], and this study further characterizes the purified protein and its interaction with microtubules. We show that purified MAP1B (...


    pub_type: 杂志文章


    authors: Pedrotti B,Ulloa L,Avila J,Islam K

    更新日期:1996-03-05 00:00:00

  • Structural and functional analysis of NADPH-cytochrome P-450 reductase from human liver: complete sequence of human enzyme and NADPH-binding sites.

    abstract::The complete amino acid sequence of human liver NADPH-cytochrome P-450 reductase has been determined by microsequence analysis and mass spectrometry. The total sequence consists of 676 amino acids initiated by an amino-terminal acetyl group. There is no evidence for posttranslational modifications, including Asn-linke...


    pub_type: 杂志文章


    authors: Haniu M,McManus ME,Birkett DJ,Lee TD,Shively JE

    更新日期:1989-10-17 00:00:00

  • Structure and assembly of the heterotrimeric and homotrimeric C-propeptides of type I collagen: significance of the alpha2(I) chain.

    abstract::Assembly of the type I procollagen molecule begins with interactions among the C-pro alpha1(I) and C-pro alpha2(I) domains. The C-propeptide domains themselves have subdomains of distinct structures. The important questions are where chain association begins and the basis of the chain selectivity which leads to the pr...


    pub_type: 杂志文章


    authors: Malone JP,Alvares K,Veis A

    更新日期:2005-11-22 00:00:00