22 research outputs found
Efeito da calcinação do resíduo de bauxita nas características reológicas e no estado endurecido de suspensões com cimento Portland
Modelling and mutational evidence identify the substrate binding site and functional elements in APC amino acid transporters
The Amino acid-Polyamine-Organocation (APC) superfamily is the main family of amino acid transporters found in all domains of life and one of the largest families of secondary transporters. Here, using a sensitive homology threading approach and modelling we show that the predicted structure of APC members is extremely similar to the crystal structures of several prokaryotic transporters belonging to evolutionary distinct protein families with different substrate specificities. All of these proteins, despite having no primary amino acid sequence similarity, share a similar structural core, consisting of two V-shaped domains of five transmembrane domains each, intertwined in an antiparallel topology. Based on this model, we reviewed available data on functional mutations in bacterial, fungal and mammalian APCs and obtained novel mutational data, which provide compelling evidence that the amino acid binding pocket is located in the vicinity of the unwound part of two broken helices, in a nearly identical position to the structures of similar transporters. Our analysis is fully supported by the evolutionary conservation and specific amino acid substitutions in the proposed substrate binding domains. Furthermore, it allows predictions concerning residues that might be crucial in determining the specificity profile of APC members. Finally, we show that two cytoplasmic loops constitute important functional elements in APCs. Our work along with different kinetic and specificity profiles of APC members in easily manipulated bacterial and fungal model systems could form a unique framework for combining genetic, in-silico and structural studies, for understanding the function of one of the most important transporter families
On the fabrication and mechanical modelling microscale bistable tensegrity systems
We report about the analysis, design, and experimental testing of modular structures composed of bistable units derived from the classic triangular tensegrity prism. Tensegrity structures are pin-connected frameworks, composed by bars and cables, possessing internal mechanisms and self-stress states, and featuring a variety of structural responses depending on their prestress, edge connectivity, and geometry. When a tensegrity system has only one internal mechanism and one self-stress state, as in the triangular prism case, it is possible to associate to it a corresponding bistable unit, by replacing all cables with bars and changing their edge-lengths slightly. After presenting experimental results of compression tests carried out on microscale specimens fabricated through multiphoton lithography, we compare them with the numerical predictions obtained by our computational model
Evaluation of Kuwaiti Argillaceous Materials as a Replacement of Sand for Clinker Production
Aspergillus nidulans CkiA is an essential casein kinase I required for delivery of amino acid transporters to the plasma membrane
Type I casein kinases are highly conserved among Eukaryotes. Of the two Aspergillus nidulans casein kinases I, CkiA is related to the δ/ε mammalian kinases and to Saccharomyces cerevisiæ Hrr25p. CkiA is essential. Three recessive ckiA mutations leading to single residue substitutions, and downregulation using a repressible promoter, result in partial loss-of-function, which leads to a pleiotropic defect in amino acid utilization and resistance to toxic amino acid analogues. These phenotypes correlate with miss-routing of the YAT plasma membrane transporters AgtA (glutamate) and PrnB (proline) to the vacuole under conditions that, in the wild type, result in their delivery to the plasma membrane. Miss-routing to the vacuole and subsequent transporter degradation results in a major deficiency in the uptake of the corresponding amino acids that underlies the inability of the mutant strains to catabolize them. Our findings may have important implications for understanding how CkiA, Hrr25p and other fungal orthologues regulate the directionality of transport at the ER-Golgi interface. © 2012 Blackwell Publishing Ltd