124 resultados para ALLOSTERIC SITE

em Queensland University of Technology - ePrints Archive


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Modulation of protein binding specificity is important for basic biology and for applied science. Here we explore how binding specificity is conveyed in PDZ (postsynaptic density protein-95/discs large/zonula occludens-1) domains, small interaction modules that recognize various proteins by binding to an extended C terminus. Our goal was to engineer variants of the Erbin PDZ domain with altered specificity for the most C-terminal position (position 0) where a Val is strongly preferred by the wild-type domain. We constructed a library of PDZ domains by randomizing residues in direct contact with position 0 and in a loop that is close to but does not contact position 0. We used phage display to select for PDZ variants that bind to 19 peptide ligands differing only at position 0. To verify that each obtained PDZ domain exhibited the correct binding specificity, we selected peptide ligands for each domain. Despite intensive efforts, we were only able to evolve Erbin PDZ domain variants with selectivity for the aliphatic C-terminal side chains Val, Ile and Leu. Interestingly, many PDZ domains with these three distinct specificities contained identical amino acids at positions that directly contact position 0 but differed in the loop that does not contact position 0. Computational modeling of the selected PDZ domains shows how slight conformational changes in the loop region propagate to the binding site and result in different binding specificities. Our results demonstrate that second-sphere residues could be crucial in determining protein binding specificity.

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Off-site Manufacture (OSM) has long been recognised, both in Australia and internationally, as offering numerous benefits to all parties in the construction process. More importantly, it is recognised as a key vehicle for driving improvement within the construction industry. The uptake of OSM in construction is however limited, despite well documented benefits. The research aims to determine the ‘state-of-the-art’ of OSM in Australia. It confirms the benefits and identifies the real and perceived barriers to the widespread adoption of OSM. Further the project identifies opportunities for future investment and research. Although numerous reports have been produced in the UK on the state of OSM adoption within that region, no prominent studies exist for the Australian context. This scoping study is an essential component upon which to build any initiatives that can take advantage of the benefits of OSM in construction. The Construction 2020 report predicted that OSM is set to increase in use over the next 5-15 years, further justifying the need for such a study. The long-term goal of this study is to contribute to the improvement of the Australian construction industry through a realisation of the potential benefits of OSM.

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Off-site manufacture (OSM) offers numerous benefits to all parties in the construction process. The uptake of OSM in Australia has, however, been limited. This limited uptake corresponds to similar trends in the UK and US, although the level of OSM there appears to be increasing. This project undertook three workshops — one each in Victoria, Queensland and Western Australia — and 18 interviews with key stakeholders to assist in identifying the general benefits and barriers to OSM uptake in the Australian construction industry. Seven case studies were also undertaken, involving construction projects that used OSM, ranging from civil projects through to residential. Each of these case studies has been analysed to identify what worked and what didn’t, and suggest the lessons to be learned from each project.

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Much has been written on Off-site Manufacture (OSM) in construction, particularly regarding the perceived benefits and barriers to implementation. However, very little understanding of the state of OSM in the Australian construction industry exists. A ‘scoping study' has recently been undertaken to determine the ‘state-of-the-art’ of OSM in Australia. This involved several industry workshops, interviews and case studies across four major states of Australia. The study surveyed a range of suppliers across the construction supply-chain, incorporating the civil, commercial and housing segments of the market. This revealed that skills shortages and lack of adequate OSM knowledge are generally the greatest issues facing OSM in Australia. The drivers and constraints that emerged from the research were, in large measure, consistent with those found in the US and UK, although some Australian anomalies are evident, such as the geographical disparity of markets. A comparative analysis with similar studies in the UK and US is reported, illustrating both the drivers and constraints confronting the industry in Australia. OSM uptake into the future is however dependent on many factors, not least of which is a better understanding of the construction process and its associated costs.

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In 2004, with the increasing overloading restriction requirements of society in Anhui, a provincial comprehensive overloading transportation survey has been developed to take evaluations on overloading actuality and enforcement efficiency with the support of the World Bank. A total of six site surveys were conducted at Hefei, Fuyang, Luan, Wuhu, Huainan and Huangshan Areas with four main contents respectively: traffic volume, axle load, freight information and registration information. Via statistical analysis on the survey data, conclusions were gained that: vehicle overloading are very universal and serious problems at arterial highways in Anhui now. The traffic loads have far exceeded the designed endure capacity of highways and have caused prevalent premature pavement damage, especially for rigid pavement. The overloading trucks are unimpeded engaged in highway freight transportation actually due to the disordered overloading enforcement strategies and the deficient inspecting technologies.

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The Internet theoretically enables marketers to personalize a Website to an individual consumer. This article examines optimal Website design from the perspective of personality trait theory and resource-matching theory. The influence of two traits relevant to Internet Web-site processing—sensation seeking and need for cognition— were studied in the context of resource matching and different levels of Web-site complexity. Data were collected at two points of time: personality-trait data and a laboratory experiment using constructed Web sites. Results reveal that (a) subjects prefer Web sites of a medium level of complexity, rather than high or low complexity; (b)high sensation seekers prefer complex visual designs, and low sensation seekers simple visual designs, both in Web sites of medium complexity; and (c) high need-for-cognition subjects evaluated Web sites with high verbal and low visual complexity more favourably.

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The interaction of quercetin, which is a bioflavonoid, with bovine serum albumin (BSA) was investigated under pseudo-physiological conditions by the application of UV–vis spectrometry, spectrofluorimetry and cyclic voltammetry (CV). These studies indicated a cooperative interaction between the quercetin–BSA complex and warfarin, which produced a ternary complex, quercetin–BSA–warfarin. It was found that both quercetin and warfarin were located in site I. However, the spectra of these three components overlapped and the chemometrics method – multivariate curve resolution-alternating least squares (MCR-ALS) was applied to resolve the spectra. The resolved spectra of quercetin–BSA and warfarin agreed well with their measured spectra, and importantly, the spectrum of the quercetin–BSA–warfarin complex was extracted. These results allowed the rationalization of the behaviour of the overlapping spectra. At lower concentrations ([warfarin] < 1 × 10−5 mol L−1), most of the site marker reacted with the quercetin–BSA, but free warfarin was present at higher concentrations. Interestingly, the ratio between quercetin–BSA and warfarin was found to be 1:2, suggesting a quercetin–BSA–(warfarin)2 complex, and the estimated equilibrium constant was 1.4 × 1011 M−2. The results suggest that at low concentrations, warfarin binds at the high-affinity sites (HAS), while low-affinity binding sites (LAS) are occupied at higher concentrations.