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Marchalonis JJ Robey I Schluter SF Yocum DE 《Applied biochemistry and biotechnology》2000,83(1-3):31-52
To characterize the binding specificity and light- and heavy-chain variable region usage in monoclonal human autoantibodies (mAAbs) to T-cell receptors, we constructed heterohybridomas from peripheral blood B cells of three rheumatoid arthritis (RA) patients. From a panel of more than 200 heterohybridomas secreting IgM autoantibodies binding to T-cell receptor Vbeta chain first complementarity determining segments (CDR1), we characterized two IgM/lambda molecules from a single patient in detail. These bound to both CDR1 peptide epitopes and intact TCR of recombinant single-chain T-cell receptor constructs, and to T-cell surface TCR. Spectratype analysis using epitopes mimicking a set of 24 Vbeta genes indicated that one molecule bound only a few members of the set, whereas the second showed considerable epitope promiscuity by binding to more than half of the tested CDR1 peptides. Both mAAbs used variants of a Vlambda3 gene that were very similar to one another and to the germline gene. The epitope-promiscuous autoantibody used a V(H)4 gene identical to a germline prototype, while the other incorporated a V(H)3 sequence differing in only a single residue from its germline prototype. The CDR3s of both were large and distinct from each other as well as from the corresponding segments of rheumatoid factors and "cold agglutinins" using the same or related V(H) germline genes. These mAAbs offer models for deciphering the basis of epitope promiscuity, and serve as candidates for direct use in immunomodulation because they are of intrinsic human origin and do not require molecular engineering to adapt them for use in therapy. 相似文献
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The minireview presented here summarizes current information on the structure and function of PsbO, the photosystem II (PSII) manganese-stabilizing protein, with an emphasis on the protein's assembly into PSII, and its function in facilitating rapid turnovers of the oxygen evolving reaction. Two putative mechanisms for functional assembly of PsbO, which behaves as an intrinsically disordered polypeptide in solution, into PSII are proposed. Finally, a model is presented for the role of PsbO in relation to the function of the Mn, Ca(2+), and Cl(-) cofactors that are required for water oxidation, as well as for the action of hydroxide and small Mn reductants that inhibit the function of the active site of the oxygen-evolving complex. 相似文献
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Aalseth CE Barbeau PS Bowden NS Cabrera-Palmer B Colaresi J Collar JI Dazeley S de Lurgio P Fast JE Fields N Greenberg CH Hossbach TW Keillor ME Kephart JD Marino MG Miley HS Miller ML Orrell JL Radford DC Reyna D Tench O Van Wechel TD Wilkerson JF Yocum KM;CoGeNT Collaboration 《Physical review letters》2011,106(13):131301
We report on several features in the energy spectrum from an ultralow-noise germanium detector operated deep underground. By implementing a new technique able to reject surface events, a number of cosmogenic peaks can be observed for the first time. We discuss an irreducible excess of bulklike events below 3 keV in ionization energy. These could be caused by unknown backgrounds, but also dark matter interactions consistent with DAMA/LIBRA. It is not yet possible to determine their origin. Improved constraints are placed on a cosmological origin for the DAMA/LIBRA effect. 相似文献
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When reaction mixtures containing 9,10-anthraquinone-2-sulfonate and chloroplasts poisoned with 3–(3,4-dichlorophenyl)-l, l′-dimethylurea were illuminated with white light, photosystem I-catalyzed cyclic photophosphorylation was observed. Illumination of identical reaction mixtures with red light produced no ATP synthesis. This phenomenon is due to photoreduction of the anthraquinone which is supported by the electron donor activity of Tricine buffer. The photoreduction reaction was used to generate reduced catalysts (anthraquinone sulfonate, menadione bisulfite) of photosystem I cyclic photophosphorylation activity. The rates of ATP synthesis obtained by this method (250–300 μmol/h-mg chlorophyll) indicate that sulfonated quinones are efficient mediators of cyclic electron transport around photosystem I. Although the activity catalyzed by these compounds is highly sensitive to dibromothymoquinone, very little decrease in activity is observed with antimycin A. 相似文献
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Santonicola MG Yocum MA Lenhoff AM Kaler EW 《Langmuir : the ACS journal of surfaces and colloids》2007,23(10):5358-5366
We study the phase behavior and microstructure of alkyl-beta-monoglucosides with intermediate chain lengths (octyl- and nonyl-beta-glucoside) in aqueous solutions containing ammonium sulfate and poly(ethylene glycol) (PEG). When the glucoside surfactants are mixed with PEG of molecular weight 3350 or larger, two different phase transitions are observed in the temperature range 0-100 degrees C, with lower and upper miscibility gaps separated by a one-phase isotropic region. Isothermal titration calorimetry is used to quantify the effect of PEG on the micellization properties of the alkyl monoglucosides, whereas small-angle neutron scattering gives insight into the microstructure of the surfactant/polymer mixtures near the liquid-liquid phase boundary. Results show that the range and the strength of the interactions in these solutions are highly affected by the presence of PEG. Solutions with nonyl-beta-glucoside contain larger micelles than those with octyl-beta-glucoside, and the intermicellar interactions are much stronger and longer ranged. The relevance of these findings for membrane protein crystallization is discussed. 相似文献
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Aalseth CE Barbeau PS Colaresi J Collar JI Diaz Leon J Fast JE Fields N Hossbach TW Keillor ME Kephart JD Knecht A Marino MG Miley HS Miller ML Orrell JL Radford DC Wilkerson JF Yocum KM;CoGeNT Collaboration 《Physical review letters》2011,107(14):141301
Fifteen months of cumulative CoGeNT data are examined for indications of an annual modulation, a predicted signature of weakly interacting massive particle (WIMP) interactions. Presently available data support the presence of a modulated component of unknown origin, with parameters prima?facie compatible with a galactic halo composed of light-mass WIMPs. Unoptimized estimators yield a statistical significance for a modulation of ~2.8σ, limited by the short exposure. 相似文献
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Hemoglobin (Hb) adducts have long been used as dosimeters of exposure to xenobiotics and endogenously formed reactive metabolites. In this study, hemoglobin chains were separated from each other and their prosthetic heme groups and reacted with 4-oxo-2-nonenal, a major breakdown product of lipid hydroperoxides. The adducts were characterized by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-TOF/MS) analysis of the intact proteins and by a combination of liquid chromatography/electrospray ionization/tandem MS (MS/MS) and MALDI-TOF/MS/MS analysis of the tryptic peptides. Covalent modifications were found on both hemoglobin chains. The location was determined to be on H20 of the alpha-hemoglobin chain and on H(63) of the beta-hemoglobin chain. Molecular modeling revealed that these two residues were two most solvent accessible H residues present in intact Hb. The proposed reaction mechanism is based on that described for the reaction of 4-hydroxy-2-nonenal with proteins. Initial nucleophilic Michael addition is followed by hydration of the resulting aldehyde, cyclization, and two sequential dehydration reactions to give stable furan derivatives. This results in the addition of 136 Da from 4-oxo-2-nonenal to give adducts corresponding to (17)VGAH(.) AGEYGAEALER(31) from alpha-hemoglobin and (62)AH(.) GK(65) from beta-hemoglobin. These hemoglobin modifications can potentially serve as biomarkers of lipid hydroperoxide-mediated macromolecule damage and may reflect an indirect measurement of the potential for DNA damage in vivo. 相似文献
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