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Interleukin-7 (IL-7) is a proteinaceous biological response modifier that has a bioactive tertiary structure dependent on disulfide bond formation. Disulfide bond assignments in human (h)IL-7 are based upon the results of matrix-assisted laser desorption/ionization (MALDI) mass spectroscopy and Cys to Ser mutational analyses. A gene encoding the hIL-7 was synthesized incorporating Escherichia coli codon usage bias and was used to express biologically active protein as determined by stimulation of precursor B-cell proliferation. MALDI mass spectroscopic analysis of trypsin-digested hIL-7 was performed and compared with the anticipated results of a simulated tryptic digestion. Many of the anticipated hIL-7 tryptic fragments were detected including one with a molecular mass equivalent to the sum of two polypeptides linked through a disulfide bond formed from Cys residues (Cys3 and Cys142). Subsequently, Cys to Ser substitution mutational analyses were performed. A hIL-7 variant with all six Cys substituted with Ser was found to be biologically inactive (EC50 > 1 x 10(-7) M). In contrast, a family of single disulfide bond-forming variants of hIL-7 were constructed by reintroducing Cys pairs (Cys3-Cys142, Cys35-Cys130, and Cys48-Cys93), and each could stimulate cell proliferation with an EC50 of 4 x 10(-9), 2 x 10(-8), and 2 x 10(-9) M, respectively. In single disulfide bond-forming mutants of hIL-7, the ability to stimulate cell proliferation was abolished in the presence of 2 mM dithiothreitol. The results presented strongly suggest that only a single disulfide bond is required for hIL-7 to form a tertiary structure capable of stimulating precursor B-cell proliferation. 相似文献
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GA Murphy MS Moore G Drivas P Pérez de la Ossa A Villamarin P D'Eustachio MG Rush 《Canadian Metallurgical Quarterly》1997,8(12):2591-2604
Ran, the small, predominantly nuclear GTPase, has been implicated in the regulation of a variety of cellular processes including cell cycle progression, nuclear-cytoplasmic trafficking of RNA and protein, nuclear structure, and DNA synthesis. It is not known whether Ran functions directly in each process or whether many of its roles may be secondary to a direct role in only one, for example, nuclear protein import. To identify biochemical links between Ran and its functional target(s), we have generated and examined the properties of a putative Ran effector mutation, T42A-Ran. T42A-Ran binds guanine nucleotides as well as wild-type Ran and responds as well as wild-type Ran to GTP or GDP exchange stimulated by the Ran-specific guanine nucleotide exchange factor, RCC1. T42A-Ran.GDP also retains the ability to bind p10/NTF2, a component of the nuclear import pathway. In contrast to wild-type Ran, T42A-Ran.GTP binds very weakly or not detectably to three proposed Ran effectors, Ran-binding protein 1 (RanBP1), Ran-binding protein 2 (RanBP2, a nucleoporin), and karyopherin beta (a component of the nuclear protein import pathway), and is not stimulated to hydrolyze bound GTP by Ran GTPase-activating protein, RanGAP1. Also in contrast to wild-type Ran, T42A-Ran does not stimulate nuclear protein import in a digitonin permeabilized cell assay and also inhibits wild-type Ran function in this system. However, the T42A mutation does not block the docking of karyophilic substrates at the nuclear pore. These properties of T42A-Ran are consistent with its classification as an effector mutant and define the exposed region of Ran containing the mutation as a probable effector loop. 相似文献
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Kelly Jeffrey A.; Murphy Debra A.; Sikkema Kathleen J.; Kalichman Seth C. 《Canadian Metallurgical Quarterly》1993,48(10):1023
Behavior change remains the only means for primary prevention of HIV disease. Psychology should take a leading role in efforts to curtail the epidemic, but has not contributed to HIV prevention at a level proportionate to the urgency of the crisis. The authors propose an updated agenda for behavioral research on AIDS-HIV prevention implementing accelerated community trials of promising behavior change models, conducting trials of community-level interventions on a large scale and focused on populations most vulnerable to HIV infections, establishing partnerships between HIV research and community service organizations, integrating efforts from across psychology disciplines to advance and refine HIV prevention interventions, and mobilizing interdisciplinary HIV prevention resources and communication mechanisms to rapidly translate research findings to community and public policy arenas. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
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A review of robust design methods for multiple responses 总被引:1,自引:0,他引:1
Terrence E. Murphy Kwok-Leung Tsui Janet K. Allen 《Research in Engineering Design》2005,16(3):118-132
Problems in engineering design often involve determining design variable settings to optimize individual product performance
for multiple criteria, which are often in conflict. We review mathematically rigorous techniques from the statistical literature
for finding a vector x of design variable settings, which produces an optimal compromise solution among a group of prioritized response variables.
The best compromise solution is typically gained by optimizing an objective function, which incorporates the prioritized demands
of multiple responses. Since most multi-response objective functions are constructed by combining the functions used to optimize
univariate responses, a review of the prominent univariate approaches is presented first. A multivariate approach from the
engineering literature called the compromise Decision Support Problem is also reviewed. Finally a table comparing the relative
merits of the different multivariate approaches summarizes the article in a concise and user-friendly fashion. 相似文献
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Lists of core competencies or critical capabilities are popular and plentiful. This article makes the case that a single core competency or critical capability accounts for professional success: the exercise of judgment. Judgment is the critical determinant of the quality of professional decisions, actions, and evaluations. Practical models of judgment are presented. The elements and dynamics of judgment are detailed. Individual, group, and organization implications are explored. Coaching and consulting interventions are explained. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
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Robert H. Doremus Denis Murphy Narottam P. Bansal William A. Lanford Chandra Burman 《Journal of Materials Science》1985,20(12):4445-4453
When liquid water contacts a zirconium-barium-lanthanum fluoride glass, at least three different processes occur. Barium and zirconium fluoride dissolve into the water, water penetrates into the glass, and zirconium fluoride crystals grow on the glass surface, in static solution. The rate of dissolution, as measured by solution analysis, is possibly controlled by diffusion in the solid surface; surface blockage and surface reactions are other possible kinetic steps involved. Diffusion in solution is not the controlling mechanism. Hydrogen profiles in the glass surface suggest that the penetration rate of water into the glass is controlled by diffusion and a surface reaction. 相似文献