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691.
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
692.
Kenji Kuwayama Kenji Tsujikawa Hajime Miyaguchi Tatsuyuki Kanamori Yuko T. Iwata Hiroyuki Inoue 《Analytical and bioanalytical chemistry》2013,405(12):3945-3952
The aim of this study was to determine whether an ingested drug and its metabolites could be detected in the subject’s fingerprints. Caffeine (CF) was chosen as the model drug. Three healthy subjects were asked to consume a cup of coffee (ca. 100 mL) containing 80 micro micro mg CF as the total dose, which is the normal amount in one cup of coffee. After washing hands with water to remove external contaminants, each subject pressed the index fingertip to a collecting matrix just before consuming the test cup of coffee, and then again pressed the index fingertip to the collecting matrix after 1, 3, 5, and 7 h. The time curve of the amounts of CF and its metabolites—theobromine (TB), paraxanthine (PX), and theophylline (TP)—in fingerprints and blood was determined using liquid chromatography/tandem mass spectrometry (LC/MS). A filter paper wetted with water (50 μL) was an efficient collecting matrix for extracting the analytes from the fingertip. With optimized sample preparation and LC/MS conditions, the total operating time, from taking the fingerprints to obtaining the analytical result, was approximately 10 min. The lower limits of quantification for CF, TB, PX, and TP were 0.5, 5, 0.5, and 5 ng/fingerprint, respectively. The amount of CF or PX determined in fingerprints obtained over 7 h after coffee intake was significantly greater than the amount determined in fingerprints taken before drinking coffee. Fingerprints were a more efficient source for drug testing than other biological samples, such as blood and sweat, because the procedures for sampling and extracting the drugs were simpler and took less time. The method could be used to prove drug intake in criminal investigations. 相似文献
693.
Hereditary Spastic Paraplegia Protein Spartin Is an FK506-Binding Protein Identified by mRNA Display
Mayuko Tokunaga Hirokazu Shiheido Ichigo Hayakawa Akiko Utsumi Hideaki Takashima Nobuhide Doi Kenichi Horisawa Yuko Sakuma-Yonemura Noriko Tabata Hiroshi Yanagawa 《Chemistry & biology》2013,20(7):935-942
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694.
Satoshi Kojima Tsugihiko Hidaka Yuko Ohba Katsuo Ohkata 《Phosphorus, sulfur, and silicon and the related elements》2013,188(3):729-732
The Horner-Wadsworth-Emmons reaction of diphenylphosphonoacetamides [(PhO) 2 P(O)CH 2 CONRR'] ( 2a : R, R' = CH 2 Ph; 2b : R = CH 2 Ph, R' = H; 2c : R = Me, R' = OMe) was examined. The reaction of 2a was found to be Z -selective for benzaldehyde with selectivities up to 94:6. Reagent 2b led to reasonable selectivity for both benzaldehyde (85:15) and 3-phenylpropionaldehyde (87:13), while 2c was somewhat effective only for the latter aldehyde (83:17). 相似文献
695.
Shunichi Aikawa Chiharu Sekiguchi Yuko Yamazaki Mika Hattori Tatsuya Isaka Yasuhiko Yoshida Shogo Ihara 《Journal of heterocyclic chemistry》2014,51(2):343-348
The reaction of N‐aryl benzamidines 1a , 1b , 1c , 1d , 1e , 1f , 1g , 1h , 1i , 1j , 1k , 1l , 1m , 1n with diphenyl carbonate 2a or ethyl phenyl carbonate 2b synthesized 2‐arylquinazolin‐4(3H)‐ones 3a , 3b , 3c , 3d , 3e , 3f , 3g , 3h , 3i , 3j , 3k , 3l , 3m , 3n in simple and safe process with good yields (71–90%). It was suggested that different electron‐donating substituent in N‐aryl benzamidines 1a , 1b , 1c , 1d , 1e , 1f , 1g , 1h , 1i , 1j , 1k , 1l , 1m , 1n afforded similar effect to the yields of 2‐arylquinazolin‐4(3H)‐ones 3a , 3b , 3c , 3d , 3e , 3f , 3g , 3h , 3i , 3j , 3k , 3l , 3m , 3n . In these reactions, N‐aryl benzamidines 1a , 1b , 1c , 1d , 1e , 1f , 1g , 1h , 1i , 1j , 1k , 1l , 1m , 1n built up intermediate compounds by nucleophilic addition to carbonates 2 to give annulation products 3a , 3b , 3c , 3d , 3e , 3f , 3g , 3h , 3i , 3j , 3k , 3l , 3m , 3n , following to cyclization involving the elimination of ethanol/phenol. 相似文献
696.
Ryoya Hiraoki Hayaka Fukuzumi Yuko Ono Tsuguyuki Saito Akira Isogai 《Cellulose (London, England)》2014,21(1):167-176
Two cellouronic acids [sodium (1 → 4)-β-polyglucuronates, CUAs] and one 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-oxidized wood cellulose (TOC) became soluble in 8 % lithium chloride/N,N-dimethylacetamide (LiCl/DMAc) after the methylation of C6 carboxyl groups in these samples using trimethylsilyldiazomethane (TMSD). The obtained solutions were diluted to 1 % LiCl/DMAc and subjected to size-exclusion chromatography combined with multi-angle laser-light scattering (SEC-MALLS). Neither depolymerization nor side reactions took place during methylation; this was confirmed by SEC-MALLS and nuclear magnetic resonance analyses, using CUAs as models. The SEC-MALLS analysis of the original wood cellulose and the carboxyl-methylated TOC prepared from it, using 1 % LiCl/N,N-dimethyl-2-imidazolidinone and 1 % LiCl/DMAc, respectively, as eluents, showed that the weight-average degree of polymerization of the original wood cellulose decreased from 3,100 to 2,210 through TEMPO-mediated oxidation. The molecular-mass distributions of the original wood cellulose and the TOC both consisted of one large peak with a small shoulder, indicating that some of the oxidized hemicelluloses remained in the TOC. The combination of methylation of carboxyl groups in polysaccharides using TMSD and subsequent SEC-MALLS analysis using 1 % LiCl/DMAc as an eluent may be applicable not only to TOCs, but also to other polysaccharides with carboxyl groups, for evaluation of their molecular-mass parameters. 相似文献
697.
698.
C5–C10 Directly Bonded Tetrodotoxin Analogues: Possible Biosynthetic Precursors of Tetrodotoxin From Newts
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Yuta Kudo Yoko Yamashita Prof. Dr. Dietrich Mebs Dr. Yuko Cho Prof. Dr. Keiichi Konoki Prof. Dr. Takeshi Yasumoto Prof. Dr. Mari Yotsu‐Yamashita 《Angewandte Chemie (International ed. in English)》2014,53(52):14546-14549
The identification of novel tetrodotoxin (TTX, 1 ) analogues would significantly contribute to the elucidation of its biosynthetic pathway. In this study, the first C5–C10 directly bonded TTX analogues, 4,9‐anhydro‐10‐hemiketal‐5‐deoxyTTX ( 2 ) and 4,9‐anhydro‐8‐epi‐10‐hemiketal‐5,6,11‐trideoxyTTX ( 3 ), were found in the newt Cynops ensicauda popei by using a screening method involving HILIC‐LC–MS/MS focused on the fragment ions of TTX analogues, and their structures were elucidated by spectroscopic methods. Compound 2 was detected in a wide range of newt species, and the 2 and TTX contents of 22 newt specimens were correlated (rs=0.88). Based on these results and its structural features, 2 was predicted to serve as a precursor of TTX that would be directly converted into 4,9‐anhydroTTX ( 4 ) by Baeyer–Villiger‐like oxidation or via 4,9‐anhydro‐5‐deoxyTTX formed by cleavage of the C5–C10 bond. The bicyclic carbon skeletons of 2 and 3 suggested a possible monoterpene origin for TTX. 相似文献
699.
Yamamoto YS Oshima Y Shinzawa H Katagiri T Matsuura Y Ozaki Y Sato H 《Analytica chimica acta》2008,619(1):8-13
A ball lens hollow-fiber Raman probe (BHRP) is a powerful tool for in vivo nondestructive subsurface analysis of biomedical tissues in a living body. It has confocal-like optical properties, but its collection volume is rather large in comparison with that of a conventional confocal Raman system. Therefore, the obtained Raman spectra have contributions from the upper and lower layers at different rates depending on the thickness of the upper layer when the measurement point is close to the boundary surface of the two layers. In the present study, we describe a methodology to extract quantitative information about the thickness of the subsurface layer structure by using a BHRP combined with the partial least-square regression (PLSR) analysis. The simulation study indicates that distribution of the collection efficiency in the collection volume of the BHRP is similar to a Gaussian distribution. The empirical study suggests that the PLSR model built with only a principal component (PC) 1 based on the linearized depth data gives good prediction. 相似文献
700.
Matsumoto S Yamaguchi S Ueno S Komatsu H Ikeda M Ishizuka K Iko Y Tabata KV Aoki H Ito S Noji H Hamachi I 《Chemistry (Weinheim an der Bergstrasse, Germany)》2008,14(13):3977-3986
In a focused library of glycolipid-based hydrogelators bearing fumaric amide as a trans-cis photoswitching module, several new photoresponsive supramolecular hydrogelators were discovered, the gel-sol/sol-gel transition of which was pseudo-reversibly induced by light. Studying the optimal hydrogel by NMR spectroscopy and various microscopy techniques showed that the trans-cis photoisomerization of the double bond of the fumaric amide unit effectively caused assembly or disassembly of the self-assembled supramolecular fibers to yield the macroscopic hydrogel or the corresponding sol, respectively. The entanglement of the supramolecular fibers produced nanomeshes, the void space of which was roughly evaluated to be 250 nm based on confocal laser scanning microscopy observations of the size-dependent Brownian motion of nanobeads embedded in the supramolecular hydrogel. It was clearly shown that such nanomeshes become a physical obstacle that captures submicro- to micrometer-sized substrates such as beads or bacteria. By exploiting the photoresponsive property of the supramolecular nanomeshes, we succeeded in off/on switching of bacterial movement and rotary motion of bead-tethered F(1)-ATPase, a biomolecular motor protein, in the supramolecular hydrogel. Furthermore, by using the photolithographic technique, gel-sol photopatterning was successfully conducted to produce sol spots within the gel matrix. The fabricated gel-sol pattern not only allowed regulation of bacterial motility in a limited area, but also off/on switching of F1-ATPase rotary motion at the single-molecule level. These results demonstrated that the photoresponsive supramolecular hydrogel and the resulting nanomeshes may provide unique biomaterials for the spatiotemporal manipulation of various biomolecules and live bacteria. 相似文献