Multicentric Carpotarsal Osteolysis Is Caused by Mutations Clustering in the Amino-Terminal Transcriptional Activation Domain of MAFB

Multicentric carpotarsal osteolysis (MCTO) is a rare skeletal dysplasia characterized by aggressive osteolysis, particularly affecting the carpal and tarsal bones, and is frequently associated with progressive renal failure. Using exome capture and next-generation sequencing in five unrelated simplex cases of MCTO, we identified previously unreported missense mutations clustering within a 51 base pair region of the single exon of MAFB, validated by Sanger sequencing. A further six unrelated simplex cases with MCTO were also heterozygous for previously unreported mutations within this same region, as were affected members of two families with autosomal-dominant MCTO. MAFB encodes a transcription factor that negatively regulates RANKL-induced osteoclastogenesis and is essential for normal renal development. Identification of this gene paves the way for development of novel therapeutic approaches for this crippling disease and provides insight into normal bone and kidney development.

Septin C-Terminal Domain Interactions: Implications for Filament Stability and Assembly

Septins form a conserved family of filament forming GTP binding proteins found in a wide range of eukaryotic cells. They share a common structural architecture consisting of an N-terminal domain, a central GTP binding domain and a C-terminal domain, which is often predicted to adopt a coiled-coil conformation, at least in part. The crystal structure of the human SEPT2/SEPT6/SEPT7 heterocomplex has revealed the importance of the GTP binding domain in filament formation, but surprisingly no electron density was observed for the C-terminal domains and their function remains obscure. The dearth of structural information concerning the C-terminal region has motivated the present study in which the putative C-terminal domains of human SEPT2, SEPT6 and SEPT7 were expressed in E. coli and purified to homogeneity. The thermal stability and secondary structure content of the domains were studied by circular dichroism spectroscopy, and homo- and hetero-interactions were investigated by size exclusion chromatography, chemical cross-linking, analytical ultracentrifugation and surface plasmon resonance. Our results show that SEPT6-C and SEPT7-C are able to form both homo- and heterodimers with a high alpha-helical content in solution. The heterodimer is elongated and considerably more stable than the homodimers, with a K (D) of 15.8 nM. On the other hand, the homodimer SEPT2-C has a much lower affinity, with a K (D) of 4 mu M, and a moderate alpha-helical content. Our findings present the first direct experimental evidence toward better understanding the biophysical properties and coiled-coil pairings of such domains and their potential role in filament assembly and stability.

On the derivation of the terminal velocity for the falling magnet from dimensional analysis

Dimensional analysis was employed to develop a predictive formula for the terminal velocity for a magnet dropped down a metallic tube. In this particular application, the technique succeeded in generating the same formula theoretically derived and that has been published by others. The analysis thus presented suggests other applications that can be developed for motivating in the use of the technique.

The role of the C-terminal region of pulchellin A-chain in the interaction with membrane model systems

Pulchellin is a Ribosome Inactivating Protein containing an A-chain (PAC), whose toxic activity requires crossing the endoplasmic reticulum (ER) membrane. In this paper, we investigate the interaction between recombinant PAC (rPAC) and Langmuir monolayers of dipalmitoyl phosphatidyl glycerol (DPPG), which served as membrane model. Three catalytically active, truncated PACs with increasing deletion of the C-terminal region, possessing 244,239 and 236 residues (rPAC(244), rPAC(239) and rPAC(236)), were studied. rPAC had the strongest interaction with the DPPG monolayer, inducing a large expansion in its surface pressure-area isotherm. The affinity to DPPG decreased with increased deletion of the C-terminal region. When the C-terminal region was deleted completely (rPAC(236)), the interaction was recovered, probably because other hydrophobic regions were exposed to the membrane. Using Polarization Modulated-Infrared Reflection Absorption Spectroscopy (PM-IRRAS) we observed that at a bare air/water interface rPAC comprised mainly alpha-helix structures, the C-terminal region had unordered structures when interacting with DPPG. For rPAC(236) the alpha-helices were preserved even in the presence of DPPG. These results confirm the importance of the C-terminal region for PAC-ER membrane interaction. The partial unfolding only with preserved C-terminal appears a key step for the protein to reach the cytosol and develop its toxic activity. (C) 2011 Elsevier B.V. All rights reserved.

Serological detection of Plasmodium vivax malaria using recombinant proteins corresponding to the 19-kDa C-terminal region of the merozoite surface protein-1

Abstract Background Serological tests to detect antibodies specific to Plasmodium vivax could be a valuable tool for epidemiological studies, for screening blood donors in areas where the malaria is not endemic and for diagnosis of infected individuals. Because P. vivax cannot be easily obtained in vitro, ELISA assays using total or semi-purified antigens are rarely used. Based on this limitation, we tested whether recombinant proteins representing the 19 kDa C-terminal region of the merozoite surface protein-1 of P. vivax (MSP119) could be useful for serological detection of malaria infection. Methods Three purified recombinant proteins produced in Escherichia coli (GST-MSP119, His6-MSP119 and His6-MSP119-PADRE) and one in Pichia pastoris (yMSP119-PADRE) were compared for their ability to bind to IgG antibodies of individuals with patent P. vivax infection. The method was tested with 200 serum samples collected from individuals living in the north of Brazil in areas endemic for malaria, 53 serum samples from individuals exposed to Plasmodium falciparum infection and 177 serum samples from individuals never exposed to malaria. Results Overall, the sensitivity of the ELISA assessed with sera from naturally infected individuals was 95%. The proportion of serum samples that reacted with recombinant proteins GST-MSP119, His6-MSP119, His6-MSP119-PADRE and yMSP119-PADRE was 90%, 93.5%, 93.5% and 93.5%, respectively. The specificity values of the ELISA determined with sera from healthy individuals and from individuals with other infectious diseases were 98.3% (GST-MSP119), 97.7% (His6-MSP119 and His6-MSP119-PADRE) or 100% (yMSP119-PADRE). Conclusions Our study demonstrated that for the Brazilian population, an ELISA using a recombinant protein of the MSP119 can be used as the basis for the development of a valuable serological assay for the detection of P. vivax malaria.

Arquitetura brutalista e estratégia de transportes no triângulo mineiro: estações ferroviárias da Mogiana e Terminal Rodoviário Presidente Castelo Branco

Em 1989, Brandão descrevia o Triângulo Mineiro como “fruto da ambiguidade de seu estigma de fazer parte de Minas, mas ser articulada economicamente a São Paulo.” A mesorregião do Triângulo Mineiro e Alto Paranaíba, faz fronteira com os estados de Goiás, São Paulo e Mato Grosso do Sul, interligando também com a Central Mineira e com o Oeste de Minas, sendo a característica de “rota de passagem” como principal fator do desenvolvimento de sua economia. O posicionamento estratégico da região, como eixo de ligação da capital paulista ao chamado Brasil Central, pode ser considerado um importante fator no estreitamento dos laços entre a região e São Paulo, somado ao sentimento de não pertencimento do Triângulo ao estado de Minas Gerais, o qual resultou por décadas em manifestações separatistas na região. A arquitetura moderna produzida no Triângulo Mineiro e Alto Paranaíba deu um salto significativo no momento de construção da nova capital federal, em finais da década de 1950, onde o papel de mediação, principalmente da cidade de Uberlândia, no processo de infra-estruturação da nova cidade foi determinante nos avanços construtivos do Triângulo. Esse momento coincidiu com o início do processo de verticalização das principais cidades da região e o aumento de arquitetos residentes nas cidades. Por meio, em especial, dos edifícios para as estações ferroviárias da Cia Mogiana, de Oswaldo Arthur Bratke em Uberaba e Uberlândia (déc. 1960) e do Terminal Rodoviário Presidente Castelo Branco em Uberlândia, dos arquitetos Fernando Graça, Flávio Almada e Ivan Curpertino (1970), este trabalho objetiva conduzir uma discussão acerca da produção de arquitetura moderna no Triângulo Mineiro ligada às estratégias de transportes intermunicipais como própria cultura de desenvolvimento econômico da região. Nos interessa valer do debate entre o uso da estética brutalista, e da própria escolha por uma arquitetura moderna, como artifício no plano de desenvolvimento das empresas de transporte, e dos governos locais. Sobretudo, discutir as interlocuções do Triângulo Mineiro com São Paulo, rebatendo-as na formação do conjunto arquitetonico moderno produzido na região. Este trabalho é fruto da pesquisa de mestrado da autora cujo tema central é a difusão da arquitetura moderna no Triângulo Mineiro e Alto Paranaíba, pelo Iau/Usp, e financiado pela Capes.