Here, we report the crystallization and structure solution of the U2AF homology motif (UHM) domain of splicing factor Puf60 fused to Escherichia coli thioredoxin A. Both modules make extensive crystallographic contacts, contributing to a well-defined crystal lattice with clear electron density for both the thioredoxin and the Puf60-UHM module. We compare two short linker sequences between the two fusion domains, GSAM and GSPPM, for which only the GSAM-linked fusion protein yielded diffracting crystals. LY2090314 purchase While specific interdomain contacts are not observed for both fusion proteins, NMR relaxation data in solution indicate reduced

interdomain mobility between the Trx and Puf60-UHM modules. The GSPPM-linked fusion protein is significantly more flexible, albeit both linker sequences have the same number of degrees of torsional freedom. Our analysis provides a rationale for the crystallization of the GSAM-linked fusion protein and indicates that in this VE-821 manufacturer case, a four-residue linker between thioredoxin A and the fused target may represent the maximal length for crystallization purposes. Our data provide an experimental basis for the rational design of linker sequences in carrier-driven crystallization and identify thioredoxin A as a powerful fusion partner that can aid crystallization of difficult targets.”
“Multiple system atrophy (MSA)

is a neurodegenerative disease involving motor abnormalities that include akinesia, rigidity and postural instability. While improved diagnostic criteria have aided the accurate diagnosis of MSA, our understanding of the neuropathological aspects underlying MSA was bolstered by the identification of a-synuclein (alpha-syn) as the primary constituent of the abnormal protein aggregates observed in the brains of MSA patients. The generation of transgenic animal models of MSA coupled with an increasing understanding of the biochemical structure and function of a-syn has highlighted a number of key pathological pathways thought to underlie the neurodegeneration

observed in MSA. This review summarizes key findings in the field, discusses current areas of debate, and describes current experimental approaches towards disease-modifying therapies.”
“An improved system for cell-free expression of protein arrays based on DNA arrays is presented. Our technology uses an array of DNA check details constructs for cell-free expression, which acts as a template instructing the generation of the corresponding protein array. Proteins are expressed locally from these templates by a cell-free transcription and translation system, and are immobilised on a separate capture surface overlayed on the DNA array. By simplifying the setup to allow protein diffusion between the slides across a gap filled with the cell-free system, we have markedly improved the evenness of the resulting protein microarrays.”
“In recent years, dihydrodipicolinate synthase (DHDPS, E. C. 4.2.1.