1,720,977 research outputs found
Protein interactions leading to conformational changes monitored by limited proteolysis: apo form and fragments of horse cytochrome c
No full textProteolysis experiments have been used to monitor the conformational transitions from an unfolded to a folded state occurring when the apo form of horse cytochrome c (cyt c) binds the heme moiety or when two fragments of cyt c form a native-like 1:1 complex. Proteinase K was used as a proteolytic probe, in view of the fact that the broad substrate specificity of this protease allows digestion at many sites along a polypeptide chain. The rather unfolded apo form of cyt c binds heme with a concomitant conformational transition to a folded species characterized by an enhanced content of helical secondary structure. While the holoprotein is fully resistant to proteolytic digestion and the apoprotein is digested to small peptides, the noncovalent complex of the apoprotein and heme exhibits an intermediate resistance to proteolysis, in agreement with the fact that the more folded structure of the complex makes the protein substrate more resistant to proteolysis. The noncovalent native-like complex of the two fragments 1-56 and 57-104 of cyt c, covering the entire polypeptide chain of 104 residues of the protein, is rather resistant to proteolysis, while the individual fragments are easily digested. Fragment 57-104 is fast degraded to several peptides, while fragment 1-56 is slowly degraded stepwise from its C-terminal end, leading initially mostly to fragments 1-48 and 1-40 and, at later stages of proteolysis, fragments 1-38, 1-35, 1-33, and 1-31. Thus, proteolysis data indicate that the heme containing fragment 1-56 has a rather compact core and a C-terminal flexible tail. Upon prolonged incubation of the complex of fragments 1-56 and 57-104 (nicked cyt c) with proteinase K, a chain segment is removed from the nicked protein, leading to a gapped protein complex of fragments of 1-48 and 57-104 and, on further digestion, fragments 1-40 and 57-104. Of interest, the chain segment being removed by proteolysis of the complex matches the omega-loop which is evolutionarily removed in cyt c of microbial origin. Overall, rates and/or resistance to proteolysis correlates well with the extent of folding of the protein substrates, as deduced from circular dichroism measurements. Thus, our results underscore the utility of proteolytic probes for analyzing conformational and dynamic features of proteins. Finally, a specific interest of the cyt c fragment system herewith investigated resides in the fact that the fragments are exactly the exon products of the cyt c gene
Complementation of protein fragments: novel systems derived from cytochrome c and apomyoglobin
No full textGoing Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Limited proteolysis of human growth hormone at low pH: isolation, characterization, and complementation of the two biologically relevant fragments 1-44 and 45-191
No full textThe limited proteolysis approach was used to analyze the conformational features of human growth hormone (hGH) under acidic solvent conditions (A-state). Pepsin was used as the proteolytic probe because of its poor substrate specificity and its activity at low pH. Limited proteolysis of hGH in its A-state results in a selective cleavage of the Phe44-Leu45 peptide bond, leading to the production of fragments 1-44 and 45-191. The two fragments were isolated in homogeneous form for studying their conformational properties by means of spectroscopic methods. Fragment 1-44 was shown to retain little secondary and tertiary structure at neutral pH, while fragment 45-191 independently folds into a highly helical secondary structure. In particular, we have shown that the two peptic fragments are able to associate into a stable and native-like hGH complex 1-44/45-191. Our proteolysis data indicate that in acid solution hGH adopts a partly folded state characterized by a local unfolding of the first minihelix (residues 38-47) encompassing the Phe44-Leu45 peptide bond. Of interest, hGH has both insulin-like and diabetogenic effects. Two fragments of hGH occur in vivo and exert these two opposite activities, namely, fragment 1-43 showing an insulin-potentiating effect and fragment 44-191 showing a diabetogenic activity. The results of this study suggest that the conformational changes of hGH induced by an acidic pH promote the generation of the two physiologically relevant fragments by proteolytic processing of the hormone. Although pepsin cannot be the enzyme responsible for the in vivo processing of the hormone, we propose that limited proteolysis of hGH at low pH is physiologically relevant, since the hormone is exposed to an acidic environment in the cell. This study reports for the first time the analysis of the conformational features of the two individual functional domains of hGH and of their complex
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Simulating Floating Head Pressure Control with Artificial Intelligence
Full text linkIn this work, we model with artificial intelligence techniques one of the most used solutions for energy saving in the field of refrigeration. This solution, called floating head pressure control, allows for the optimum pressure condensation depending on the working environment, thus increasing the overall efficiency of the plant. Usually, these mechanisms are controlled by algorithms that are stored in the control memory of the chillers. Under certain environmental conditions, the optimal temperature and condensation pressure fluctuate with the environmental temperature. Thus, the absorbed electricity depends on the working parameters fixed by the control system of the chiller. Results in energy savings in these terms could grow as the external temperature decreases. The problem addressed in this work regards standard floating head pressure control (FHPC) systems that do not combine working environmental factors with the conditions of the machine and historical data because they are implemented adopting static models. We provide evidence about the potential of machine learning models in predicting the velocity of fans in order to adjust their load based on environmental factors and the conditions of the machine. A good setting of fan velocity will result in lower energy consumption. To do that we analyzed and implemented machine learning algorithms to provide instruments that support the operation of the chiller, enabling the floating head pressure control mode in the control system of the machine, during its process. We performed an empirical evaluation on both synthetic and real data to assess the quality of our proposal. Synthetic data are produced by an industrial software that simulates the behavior of chillers, while real-world data is collected from a commercial chiller. The results show that machine learning approaches are able to approximate real data getting errors that are 5 times smaller than the errors committed by the system that is now adopted. All the source code as well as the datasets will be made available online after the review process
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