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61.
62.
桑蚕丝素蛋白初始结构对其矿化作用的影响 总被引:4,自引:0,他引:4
以碱金属离子诱导桑蚕丝素蛋白溶液发生构象转变, 研究了蛋白质初始结构对其矿化作用的影响. FT-IR, XRD和SEM等测试结果显示, 未经任何处理的桑蚕丝素蛋白溶液矿化后形成片状复合物, 其无机相以二水磷酸氢钙(DCPD)为主; 而经过K+和Na+金属离子处理后, 桑蚕丝素溶液的结构由无规线团/螺旋构象向β-折叠发生转变, 矿化后成纤维状, 并相互结合呈现纳米级的三维多孔结构, 其无机相以热力学稳定的羟基磷灰石(HA)为主. 可以认为, 丝素蛋白结构转化为较伸展的β-折叠后, 使得更多的亲水基团暴露在外面, 在丝素蛋白分子不断凝聚成纤过程中, HA结晶快速生长并附着在这些微纤上, 最终形成纤维状的丝素蛋白/HA复合物. 该结果为阐明蛋白质的生物矿化过程及其调控机理提供了理论依据, 同时可以从矿化复合物的形成来反映这些微量元素可能对骨组织形成的影响, 为临床骨组织的修复提供一定的参考. 相似文献
63.
ZNF191 (243-368), a new human zinc finger protein, probably relates to some hereditary diseases and cancers, To obtain adequate amount of ZNF191(243-368) for the study of its property, structure and function, three different expression systems of inclusion-body, glutathione S-transferase (GST), and hexahistidine (6 × His) were used and compared. Among these systems, the expression level of ZNFI91(243-368) was increased in inclusion body system under a higher isopropylthio-β-D-galactoside (IPTG) concentration, but the non-target proteins were also increased more, which made its purification more difficult and the yield lower. The expression of His-tag fusion protein was almost not affected by IPTG concentration, temperature and inducing time. At a high IPTG concentration the highest expression yield for GST fusion protein was obtained. And the fusion proteins can be partially purified by a single affinity chromatography step. The fusion protein systems show advantages for expression of these proteins. 相似文献
64.
Josef Janča 《Mikrochimica acta》1994,112(5-6):197-215
The primary field forces can generate spatially oriented gradient of the effective property of a continuum or pseudo-continuum fluid (carrier liquid). When this gradient is coupled with the action of a secondary field of identical or different nature the isoperichoric focused zones of the dispersed species can appear. Consequently, they can be separated according to differences responding to the property gradient of the carrier liquid. This concept can be applied under static (non-flow) conditions in thin layer focusing as well as under dynamic conditions with the elution due to the carrier liquid flow in focusing field-flow fractionation. The gradient established by the action of the primary field and the concentration distribution of the isoperichoric focused zone formed by the coupled effect of the gradient and of the primary or secondary field are described theoretically. The rigorous relationship describing the shape of the focused zone is compared with the approximate solutions. The performances of the proposed principle were evaluated by model calculations. Potential experimental configurations considering the implementation of the static and dynamic conditions are discussed. The generalized isoperichoric focusing theory can be applied to describe the particular processes operating in analytical and preparative focusing separations of the particles of various, but especially of biological origin. 相似文献
65.
66.
High-level expression of soluble human β-defensin-2 fused with green fluorescent protein in Escherichia coli cell-free system 总被引:2,自引:0,他引:2
Human β-defensin-2 (hBD2), a small cationic peptide, exhibits a broad range of antimicrobial activity and does not acquire
any microbial resistance. To produce this uneasily detectable, degradable, and toxic polypeptide efficiently, an alternative
approach based on the Escherichia coli cell-free biosynthesis system was proposed. The approach implies that a polypeptide of interest is synthesized as a fusion
protein linked to a green fluorescent protein (GFP) through a cleavable spacer. With batch-mode operation, a significant amount
of hBD2 fused with GFP (0.25 mg/mL) can be expressed in this cell-free system. The productivity of the fusion protein can
be improved up to 1.2 mg/mL by employing a continuous-exchange cell-free system. Furthermore, the GFP moiety provides directly
visible and quantitative monitoring of the polypeptide synthesis, and the product is soluble and stable. This work will be
helpful in allowing the rapid and visible expression of other similar defensins using an in vitro cell-free system. 相似文献
67.
Young-ah WooChang-hee Cho Hyo-jin Kim Jong-soo YangKi-yong Seong 《Microchemical Journal》2002,73(3):299-306
A rapid and nondestructive near infrared (NIR) method using soft independent modeling of class analogy (SIMCA) for the classification of cultivation area (Korea and China) was evaluated and confirmed. Raw, first, and second derivative NIR spectra were compared to develop a robust classification rule. The chemical properties of ginseng samples were also investigated to find out the differences between Korean samples and Chinese samples. These differences make NIR spectroscopic method viable. The average value of each Korean and Chinese ginseng sample for crude fiber, crude protein, starch, and 10 inorganic constituents were measured and compared with F-test and t-test. The inorganic constituents were also measured by induced coupled plasma-atomic emission spectroscopy (ICP-AES). It could be found that the amount of starch and ten inorganic elements for example Na, Mg, P, K, Ca, Mn, Fe, Ni, Cu and Zn in ginseng samples are considerably different based on cultivation area. SIMCA has been applied to the inorganic data to investigate the possibility of ICP-AES as classification tool. However, it was observed that the result was not equal to than NIR spectra data. The overall results showed the availability of NIR method using SIMCA would be adequate for classification of cultivation of ginseng, since NIR spectra includes useful and various information on chemical properties in spite of broad and overlapped bands. 相似文献
68.
《Electroanalysis》2005,17(20):1861-1864
Electrochemical study of barley grain lipid‐transfer protein (LTP) revealed that it may belong to the metal‐binding protein class. Using differential pulse polarography the presence of Cu(II) and Zn(II) ions in the native LTP structure was proved, as well as its affinity for binding Ni(II) ion. Application of a more sensitive electroanalytical technique, such as anodic stripping voltammetry with analyte preconcentration, revealed the presence of Pb(II) and Cd(II) ions and also enabled the following Hg(II) ion binding. Possible biological role of LTP in plant stress response and its contribution to barley phytoextraction potential are discussed. 相似文献
69.
Analytical procedures for quantification of peptides in pharmaceutical research by liquid chromatography–mass spectrometry 总被引:6,自引:0,他引:6
John H Walden M Schäfer S Genz S Forssmann WG 《Analytical and bioanalytical chemistry》2004,378(4):883-897
Peptide quantification by liquid chromatography–mass spectrometry (LC–MS) combines the high resolving power of reversed-phase (RP) chromatography with the excellent selectivity and sensitivity of mass spectrometric detection. On the basis of comprehensive practical experience in the analysis of small molecules, pharmaceutical research is developing technologies for analysis of a growing number of peptidic drug candidates. This article is a detailed review of procedures based on LC–MS techniques for quantitative determination of peptides. With the focus on pharmaceutical applications several technologies for sample preparation, various aspects of peptide chromatography, important characteristics of ESI–MS, selectivity of MS-detection modes, the large variability of internal standards, and modern instrumentation are discussed. The demand for reliable, robust, sensitive, and accurate methods is discussed using numerous examples from the literature, complemented by experiments and results from our laboratory. 相似文献
70.
Brunsveld L Watzke A Durek T Alexandrov K Goody RS Waldmann H 《Chemistry (Weinheim an der Bergstrasse, Germany)》2005,11(9):2756-2772
Prenylated proteins with non-native functionalities are generally very difficult to obtain by recombinant or enzymatic means. The semisynthesis of preparative amounts of prenylated Rab guanosine triphosphatases (GTPases) from recombinant proteins and synthetic prenylated peptides depends largely on the availability of functionalised prenylated peptides corresponding to the proteins' native structure or modifications thereof. Here, we describe and compare solution-phase and solid-phase strategies for the generation of peptides corresponding to the prenylated C terminus of Rab7 GTPase. The solid-phase with utilisation of a hydrazide linker emerges as the more favourable approach. It allows a fast and practical synthesis of pure peptides and gives a high degree of flexibility in their modification. To facilitate the analysis of semisynthetic proteins, the synthesised peptides were equipped with a fluorescent group. Using the described approach, we introduced fluorophores at several different positions of the Rab7 C terminus. The position of the incorporated fluorescent groups in the peptides did not influence the protein-ligation reaction, as the generated peptides could be ligated onto thioester-tagged Rab7. However, it was found that the positioning of the fluorescent group had an influence on the functionality of the Rab7 proteins; analysis of the interaction of the semisynthetic Rab7 proteins with REP (Rab escort protein) and GDI (guanosine diphosphate dissociation inhibitor) molecules revealed that modification of the peptide side chains or of the C-terminal isoprenoid did not significantly interfere with complex formation. However, functionalisation of the C terminus was found to have an adverse effect on complex formation and stability, possibly reflecting low structural flexibility of the Rab GDI/REP molecules in the vicinity of the lipid-binding site. 相似文献