Human regulatory protein Ki-1/57 has characteristics of an intrinsically unstructured protein

The human protein Ki-1/57 was first identified through the cross reactivity of the anti-CD30 monoclonal antibody Ki-1; in Hodgkin lymphoma cells. The expression of Ki-1/57 in diverse cancer cells and its phosphorylation in peripheral blood leukocytes after mitogenic activation suggested its possible role in cell signaling. Ki-1/57 interacts with several other regulatory proteins involved in cellular signaling, transcriptional regulation and RNA metabolism, suggesting it may have pleiotropic functions. In a previous spectroscopic analysis, we observed a low content of secondary structure for Ki-1/57 constructs. Here, Circular dichroism experiments, in vitro RNA binding analysis, and limited proteolysis assays of recombinant Ki-1/57(122-413) and proteolysis assays of endogenous full length protein from human HEK293 cells suggested that Ki-1/57 has characteristics of an intrinsically unstructured protein. Small-angle X-ray scattering (SAXS) experiments were performed with the C-terminal fragment Ki-1/57(122-413). These results indicated an elongated shape and a partially unstructured conformation of the molecule in solution, confirming the characteristics of an intrinsically unstructured protein. Experimental curves together with ab initio modeling approaches revealed an extended and flexible molecule in solution. An elongated shape was also observed by analytical gel filtration. Furthermore, sedimentation velocity analysis suggested that Ki-1/57 is a highly asymmetric protein. These findings may explain the functional plasticity of Ki-1/57, as suggested by the wide array of proteins with which it is capable of interacting in yeast two-hybrid interaction assays.

Structural basis for branching-enzyme activity of glycoside hydrolase family 57: Structure and stability studies of a novel branching enzyme from the hyperthermophilic archaeon Thermococcus Kodakaraensis KOD1

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Desempenho e morfometria duodenal de frangos de corte submetidos a diferentes níveis de energia e programas de alimentação de 42 a 57 dias de idade

This experiment was conducted to evaluate the use of different energy levels and amino acid recommendations on performance, carcass yield and intestinal morphometry of broilers from 42 to 57 days of age. We used 1,600 one-day old male broilers (Cobb 500) in a completely randomized design arranged in a 2x4 factorial scheme with 3,200 and 3,600 kcal ME/kg and four different feed programs. The metabolizable energy levels and the feed programs did not determine significant differences in carcass characteristics for choosing a level of energy or a feed program, thus the energy level and the recommendations of amino acids that determine the highest cost-benefit ratio should prevail. It was observed that 3,600 kcal ME/kg resulted in some improvement on performance and morphometry of the intestinal mucosa, also the fractioning of digestible amino acid requirements during two periods resulted in worse performance and intestinal villi height.