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461.
Capon DJ Kaneko N Yoshimori T Shimada T Wurm FM Hwang PK Tong X Adams SA Simmons G Sato TA Tanaka K 《Proceedings of the Japan Academy. Series B, Physical and biological sciences》2011,87(9):603-616
There is a significant need for antibodies that can bind targets with greater affinity. Here we describe a novel strategy employing chemical semisynthesis to produce symmetroadhesins: antibody-like molecules having nonprotein hinge regions that are more flexible and extendible and are capable of two-handed binding. Native chemical ligation was carried out under mild, non-denaturing conditions to join a ligand binding domain (Aβ peptide) to an IgG1 Fc dimer via discrete oxyethylene oligomers of various lengths. Two-handed Aβ-Fc fusion proteins were obtained in quantitative yield and shown by surface plasmon resonance to bind an anti-Aβ antibody with a K(D) at least two orders of magnitude greater than the cognate Aβ peptide. MALDI-TOF MS analysis confirmed the protein/nonprotein/protein structure of the two-handed molecules, demonstrating its power to characterize complex protein-nonprotein hybrids by virtue of desorption/ionization mediated by peptide sequences contained therein. We anticipate many applications for symmetroadhesins that combine the target specificity of antibodies with the novel physical, chemical and biological properties of nonprotein hinges. 相似文献
462.
Makoto Watanabe Kazuya Terasawa Kaoru Kaneshiro Hiromasa Uchimura Rie Yamamoto Yuko Fukuyama Kazuharu Shimizu Taka-Aki Sato Koichi Tanaka 《Analytical and bioanalytical chemistry》2013,405(12):4289-4293
Protein glycosylation analysis is important for elucidating protein function and molecular mechanisms in various biological processes. We previously developed a glycan analysis method using a 3-aminoquinoline/α-cyano-4-hydroxycinnamic acid liquid matrix (3-AQ/CHCA LM) and applied it to the quantitative glycan profiling of glycoproteins. However, information concerning glycosylation sites is lost; glycopeptide analysis is therefore required to identify the glycosylation sites in glycoproteins. Human epidermal growth factor receptor 2 (HER2) is a glycoprotein that plays a role in the regulation of cell proliferation, differentiation, and migration. Several reports have described the structure of HER2, but the structures of N-glycans attached to this protein remain to be fully elucidated. In this study, 3-AQ/CHCA LM was applied to tryptic digests of HER2 to reveal its N-glycosylation state and to evaluate the utility of this LM in characterizing glycopeptides. Peptide sequence coverage was considerably improved compared to analysis of HER2 using α-cyano-4-hydroxycinnamic acid or 2,5-dihydroxybenzoic acid. Most of the peaks observed using only this LM were localized at the inner or outer regions of sample spots. Furthermore, five of the peptide peaks that were enriched within the inner region were confirmed to be glycosylated by MS/MS analysis. Three glycosylation sites were identified and their glycan structures were elucidated. The reduction in sample complexity by on-target separation allowed for higher sequence coverage, resulting in effective detection and characterization of glycopeptides. In conclusion, these results demonstrate that MS-based glycoprotein analysis using 3-AQ/CHCA is an effective method to identify glycosylation sites in proteins and to elucidate the glycan structures of glycoproteins in complex samples. 相似文献
463.
Abstract The cationic polymerization of β-phenylvinyl alkyl ethers (alkyl: methyl, ethyl, n-propyl, and n-butyl) was examined in toluene and in methylene chloride with boron fluoride etherate and with stannic chloride at ?78°C. Despite the fact that these monomers have bulky substituents on the α –and β-carbons, they could be homopoly-merized easily even in nonpolar sovent at ?78°C. The general features of β-phenylvinyl alkyl ethers in homopolymerization and co-polymerization with n-butyl vinyl ether suggested that β-phenylvinyl alkyl ethers behaved as derivatives of vinyl ethers, not as derivatives of styrenes. The polymerization products were white powders having a high softening point. 相似文献
464.
Yamaguchi M Nakayama D Shima K Kuyama H Ando E Okamura TA Ueyama N Nakazawa T Norioka S Nishimura O Tsunasawa S 《Rapid communications in mass spectrometry : RCM》2008,22(20):3313-3319
We have developed a new method to determine the N-terminal amino acid sequences of proteins, regardless of whether their N-termini are modified. This method consists of the following five steps: (1) reduction, S-alkylation and guanidination for targeted proteins; (2) coupling of sulfo-NHS-SS-biotin to N(alpha)-amino groups of proteins; (3) digestion of the modified proteins by an appropriate protease followed by oxidation with performic acid; (4) specific isolation of N-terminal peptides from digests using DITC resins; (5) de novo sequence analysis of the N-terminal peptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) using the CAF (chemically assisted fragmentation) method or tandem mass spectrometric (MS/MS) analysis according to unblocked or blocked peptides, respectively. By employing DITC resins instead of avidin resins used in our previous method (Yamaguchi et al., Rapid Commun. Mass Spectrom. 2007; 21: 3329), it has been possible to isolate selectively N-terminal peptides from proteins regardless of modification of N-terminal amino acids. Here we propose a universal method for N-terminal sequence analysis of proteins. 相似文献
465.
Huang YQ Shen ZL Okamura TA Wang Y Wang XF Sun WY Yu JQ Ueyama N 《Dalton transactions (Cambridge, England : 2003)》2008,(2):204-213
Seven new silver(I) complexes of the formula [Ag2(L)2(CF3SO3)2] (1), [Ag2(L)2(CH3SO3)2] (2) [Ag2(L)2](BF4)2 (3), [Ag3(L)2(NO3)2]NO3.5H2O (4), [Ag2(L)(NO3)2].CH3OH (5), [Ag2(L)2](ClO4)2 (6) and [Ag3(L)2(CH3CN)3](ClO4)3 (7) have been synthesized by reactions of 1,3,5-tris(2-oxazolinyl)benzene (L) with varied silver(I) salts under different conditions. The influences of counter anions and reaction conditions on the structure of the complexes are discussed. Three complexes , 1, 2 and 3 with two kinds of different 1D chain structures were obtained under the same synthetic conditions by using different silver(I) salts, and the ligand L was found to adopt bis-monodentate (1 and 2) and tris-monodentate (3) coordination modes respectively. On the other hand, by using the same silver(I) nitrate or silver(I) perchlorate but different reaction solvents, 4 and 5 or 6 and 7 were isolated respectively. Complexes 4and 5 have different 1D chain structures, and 6 is isostructural with . However, 7 is a tri-nuclear, propeller-shaped M3L2 supramolecular capsule in which L adopts a cis,cis,cis-conformation, while the ligand L in 3-6 has cis,trans,trans-conformation. The results revealed that the nature of the counter anions, such as their size, coordination ability and coordination mode, and the reaction conditions all have great impact on the structure of the complexes. The complexes were also characterized by electrospray mass spectrometry. Furthermore, complex 7 exhibited modest second-harmonic-generation (SHG) efficiency. 相似文献
466.
Giordani C Wakai C Okamura E Matubayasi N Nakahara M 《The journal of physical chemistry. B》2006,110(31):15205-15211
Self-diffusion coefficients (D) are measured for normal (nondeuterated) and deuterated cholesterol-d(6) (C26 and C27 methyl groups deuterated) in 1-octanol, chloroform, and cyclohexane at concentrations of 1-700 mM by varying the impurity water concentration (>2 mM) and temperature (30-50 degrees C). The pulsed field gradient spin-echo (PGSE) (1)H and (2)H NMR were used, respectively, at 600 and 92 MHz. At 30 degrees C, the hydrodynamic radius (R) obtained at 20 mM from the D value and solvent viscosity is 5.09, 7.07, and 6.17 A, respectively, in 1-octanol, chloroform, and cyclohexane when the impurity water is negligible. The R value in 1-octanol is the smallest and comparable with the average length of the molecular axes for the cholesterol molecule. In 1-octanol, R is invariant against the concentration variation, whereas in chloroform, R is larger and increases almost linearly with cholesterol concentration. At the highest concentration, 700 mM, the R in chloroform is 13.5 and 16.7 A, respectively, when the impurity water is at negligible and saturated concentrations. The R value larger than that in hydrogen-bonding 1-octanol indicates that cholesterol forms an aggregate through hydrogen bonding. The aggregate structure is confirmed by comparing NOESY spectra in chloroform and 1-octanol. The NOESY analysis reveals the presence of one extra cross peak (C4-C19) in chloroform compared to 1-octanol. Because the carbon atoms related to the cross peak are close to the hydroxyl group (C3-OH), cholesterol molecules are considered to be not piled but are found to be OH-centered in the aggregate. This is supported also by larger rotational hydrodynamic radii measured on cholesterol deuterated at positions C2, C3, C4, and C6. This shows that the aggregate formation is driven by the hydrogen-bonding between cholesterol molecules. 相似文献
467.
We describe a highly efficient template-directed photoligation of oligodeoxynucleotides (ODNs) through 5-carbamoylvinyl-2'-deoxycytidine ((CV)C). When an ODN containing (CV)C at the 5' end was photoirradiated with an ODN containing a pyrimidine base at the 3' end in the presence of template DNA, efficient photoligation was observed without any byproduct formation. Single nucleotide differences can be successfully distinguished by using photoligation-based DNA chip assay. [structure: see text] 相似文献
468.
Kazuhiro Kobayashi Yuta Okamura Shuhei Fukamachi Hisatoshi Konishi 《Tetrahedron》2010,66(40):7961-7964
A facile method for the synthesis of 3-substituted 3H-indol-3-ols has been developed. Thus, 2-isocyanophenyl ketones are allowed to react with various Grignard reagents to give the corresponding desired indolol derivatives in generally fair to good yields. The formation of 3-aryl-2,3-dimethylindolin-3-ols by the reaction of 2-isocyanobenzophenones with 2 M amounts of methylmagnesium bromide is also reported. 相似文献
469.
Jing Xu Qing Liu Taka-aki Okamura Man-Sheng Chen Wei-Yin Sun Norikazu Ueyama 《Solid State Sciences》2009,11(11):1903-1907
Reactions of ligand 5-(1H-imidazol-4-yl)methylaminoisophthalic acid (H3L) with varied lanthanide metal salts led to the formation of five scalelike 2D layered complexes {[Ln(H2L)(HL)(H2O)2]·H2O}n [Ln(III) = Pr(III) (1), Nd(III) (2), Sm(III) (3), Gd(III) (4), Tb(III) (5)]. The single crystal X-ray diffraction analyses revealed that five complexes crystallized in the same monoclinic space group C2/c are isomorphous and isostructural, and the 2D networks are further connected by hydrogen bonds and π–π interactions resulting in formation of 3D structures. Investigations on the visible luminescent property of the complexes demonstrate that compounds 3 and 5 show characteristic emissions of Sm(III) and Tb(III) in the solid state at room temperature, respectively. 相似文献
470.
Yuka Ikemoto Michio Ishikawa Satoru Nakashima Hidekazu Okamura Yuichi Haruyama Shinji Matsui Taro Moriwaki Toyohiko Kinoshita 《Optics Communications》2012,285(8):2212-2217
We report the status of a scattering near-field microspectroscopy apparatus developed at SPring-8 using an infrared synchrotron radiation (IR-SR) source. It consists of a scattering type scanning near-field optical microscope and a Fourier transform infrared spectrometer. The IR-SR is used as a highly brilliant and broad-band IR source. This apparatus has potential for application in near-field spectroscopy with high spatial resolution beyond the diffraction limit. In order to eliminate background scatterings from the probe shaft and/or sample surface, we used higher harmonic demodulation method. The near-field spectra were observed by 2nd harmonic components using the lock-in detection. The spatial resolution of about 300 nm was achieved at around 1000 cm? 1 (10 μm wavelength). 相似文献