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排序方式: 共有313条查询结果,搜索用时 15 毫秒
1.
Silica nanoparticles for separation of biologically active amines by capillary electrophoresis with laser-induced native fluorescence detection 总被引:1,自引:0,他引:1
This paper describes the analysis of biologically active amines by capillary electrophoresis (CE) in conjunction with laser-induced native fluorescence detection. In order to simultaneously analyze amines and acids as well as to achieve high sensitivity, 10 mM formic acid solutions (pH < 4.0) containing silica nanoparticles (SiNPs) were chosen as the background electrolytes. With increasing SiNP concentration, the migration times for seven analytes decrease as a result of increase in electroosmotic flow (EOF) and decrease in their electrophoretic mobilities against EOF. A small EOF generated at pH 3.0 reveals adsorption of SiNPs on the deactivated capillary wall. The decreases in electrophoretic mobilities with increasing SiNP concentration up to 0.3x indicate the interactions between the analytes and the SiNPs. Having a great sensitivity (the limits of detection at a signal-to-noise ratio (S/N) = 3 of 0.09 nM for tryptamine (TA)), high efficiency, and excellent reproducibility (less than 2.4% of the migration times), this developed method has been applied to the analysis of urinal samples with the concentrations of 0.50 +/- 0.02 microM, 0.49 +/- 0.04 microM, and 74 +/- 2 microM for TA, 5-hydroxytryptamine, and tryptophan, respectively. The successful examples demonstrated in this study open up a possibility of using functional nanoparticles for the separation of different analytes by CE. 相似文献
2.
Sorel Tchewonpi Sagu Eva Landgrber Michal Rackiewicz Gerd Huschek Harshadrai Rawel 《Molecules (Basel, Switzerland)》2020,25(24)
Sorghum is of growing interest and considered as a safe food for wheat related disorders. Besides the gluten, α-amylase/trypsin-inhibitors (ATIs) have been identified as probable candidates for these disorders. Several studies focused on wheat-ATIs although there is still a lack of data referring to the relative abundance of sorghum-ATIs. The objective of this work was therefore to contribute to the characterization of sorghum ATI profiles by targeted proteomics tools. Fifteen sorghum cultivars from different regions were investigated with raw proteins ranging from 7.9 to 17.0 g/100 g. Ammonium bicarbonate buffer in combination with urea was applied for protein extraction, with concentration from 0.588 ± 0.047 to 4.140 ± 0.066 mg/mL. Corresponding electrophoresis data showed different protein profiles. UniProtKB data base research reveals two sorghum ATIs, and P81367; both reviewed and a targeted LC–MS/MS method was developed to analyze these. Quantifier peptides ELAAVPSR ( P81368) and TYMVR ( P81367) were identified and retained as biomarkers for relative quantification. Different reducing and alkylating agents were assessed and combination of tris (2 carboxyethyl) phosphine/iodoacetamide gave the best response. Linearity was demonstrated for the quantifier peptides with standard recovery between 92.2 and 107.6%. Nine sorghum cultivars presented up to 60 times lower ATI contents as compared to wheat samples. This data suggests that sorghum can effectively be considered as a good alternative to wheat. P81368相似文献
3.
In our experiment, three groups of seedlings of SDP Pharbitis nil cv. violet were sepa-rately treated with three different photoperiods (1,16 h dark period--SD; 2, continuous illumi-nation--CL; 3, 16 h dark treatment with 10 min white light in the middle of the dark period--NB). By analysing proteins in the cotyledons from three groups with 2-D PAGE, we found nodifference in protein pattern between the three groups at 0 or 8 h after photoperiodic treatments.At 24 h after the treatments, a specific protein(MW:19 kD; pI: 4.5)appeared only in the cotyledonsof the seedlings which endured SD. This protein disappeared at 72 h after SD. ActinomycinD could inhibit flowering and the specific protein occurrence when applied to cotyledonsprior to SD, but it had no inhibition effect on flowering as well as the specific proteinoccurrence when applied to cotyledons after SD. Chloroamphenicol, a protein synthesisinhibitor, inhibited flowering when applied to cotyledons prior to or immediately after SD,but it did not i 相似文献
4.
5.
Asia Al-jabiry Dr. Martin Palmer Dr. James Langridge Dr. Jeddidiah Bellamy-Carter Dr. David Robinson Prof. Neil J. Oldham 《Chemistry (Weinheim an der Bergstrasse, Germany)》2021,27(55):13783-13792
Native mass spectrometry is now an important tool in structural biology. Thus, the nature of higher protein structure in the vacuum of the mass spectrometer is an area of significant interest. One of the major goals in the study of gas-phase protein structure is to elucidate the stabilising role of interactions at the level of individual amino acid residues. A strategy combining protein chemical modification together with collision induced unfolding (CIU) was developed and employed to probe the structure of compact protein ions produced by native electrospray ionisation. Tractable chemical modification was used to alter the properties of amino acid residues, and ion mobility-mass spectrometry (IM-MS) utilised to monitor the extent of unfolding as a function of modification. From these data the importance of specific intramolecular interactions for the stability of compact gas-phase protein structure can be inferred. Using this approach, and aided by molecular dynamics simulations, an important stabilising interaction between K6 and H68 in the protein ubiquitin was identified, as was a contact between the N-terminus and E22 in a ubiquitin binding protein UBA2. 相似文献
6.
《Angewandte Chemie (International ed. in English)》2017,56(36):10656-10669
After the discovery of insulin as a drug for diabetes, the pharmaceutical companies were faced with the challenge to meet the demand for insulin with the highest possible degree of purity in the required quantities from animal sources. The observation of an immune reaction of patients to insulin from animal pancreatic extracts made the availability of human insulin of highest priority. Only the enzyme‐catalyzed semisynthesis at the C‐terminus of the insulin B‐chain led to a commercial process, but it depended on porcine insulin and was aggravated by supply concerns. The advent of rDNA technology allowed the commercial preparation of human insulin by biosynthesis in virtually unlimited quantities. An increased chemical diversity was only envisaged through chemical synthesis, which was simplified by advances in solid‐phase peptide synthesis and chemical ligation. Single‐chain insulin precursors are now being synthesized that should enable fast screening of insulin analogues for improved biophysical, biological, and thus promising new therapeutic properties, as well as for the industrial manufacture of insulin analogues not accessible by biosynthesis. 相似文献
7.
Yan-Yan Si Lu-Jun Liang Shan Tang Yun-Kun Qi Yong Huang Ji-Shen Zheng 《Tetrahedron letters》2018,59(3):268-271
Protein chemical synthesis can provide the homogeneous atypical ubiquitin chains that are usually difficult to obtain by other methods. Herein, we report a new one-pot ligation approach for the synthesis of atypical Ub chains based on the Tfacm-protected isoUb building block with operational simplicity and high efficiency. The key intermediate, the Tfacm-protected isoUb building block can be readily prepared by Fmoc SPPS. The practicality and efficiency of the new method is demonstrated by the successful preparation of K11-linked di-Ub. 相似文献
8.
Mao Chen Ting Zhao Zhong‐Xin Zhu Chao Niu Dan‐dan Ruan Mao‐Wei Ni Xuan Zhou Li‐Tai Jin 《Electrophoresis》2013,34(24):3277-3286
An improved Stains‐All (ISA) staining method for phosphoproteins in SDS‐PAGE was described. Down to 0.5–1 ng phosphoproteins (α‐casein, β‐casein, or phosvitin) can be successfully selectively detected by ISA stain, which is approximately 120‐fold higher than that of original Stains‐All stain, but is similar to that of commonly used Pro‐Q Diamond stain. Furthermore, unlike the original Stains‐All protocol that was time consuming and light unstable, ISA stain could be completed within 60 min without resorting to protect the gels from light during the whole staining procedure. According to the results, it is concluded that ISA stain is a rapid, sensitive, specific, and economic staining method for a broad application to the research of phosphoproteins. 相似文献
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10.
Dr. Christina I. Schroeder Dr. Lachlan D. Rash Xavier Vila‐Farrés Dr. K. Johan Rosengren Dr. Mehdi Mobli Prof. Glenn F. King Prof. Paul F. Alewood Prof. David J. Craik Dr. Thomas Durek 《Angewandte Chemie (International ed. in English)》2014,53(4):1017-1020
Mambalgins are a novel class of snake venom components that exert potent analgesic effects mediated through the inhibition of acid‐sensing ion channels (ASICs). The 57‐residue polypeptide mambalgin‐2 (Ma‐2) was synthesized by using a combination of solid‐phase peptide synthesis and native chemical ligation. The structure of the synthetic toxin, determined using homonuclear NMR, revealed an unusual three‐finger toxin fold reminiscent of functionally unrelated snake toxins. Electrophysiological analysis of Ma‐2 on wild‐type and mutant ASIC1a receptors allowed us to identify α‐helix 5, which borders on the functionally critical acidic pocket of the channel, as a major part of the Ma‐2 binding site. This region is also crucial for the interaction of ASIC1a with the spider toxin PcTx1, thus suggesting that the binding sites for these toxins substantially overlap. This work lays the foundation for structure–activity relationship (SAR) studies and further development of this promising analgesic peptide. 相似文献