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This study describes the further extension of the resonant recognition model for the analysis and prediction of protein--protein and protein--DNA structure/function dependencies. The model is based on the significant correlation between spectra of numerical presentations of the amino acid or nucleotide sequences of proteins and their coded biological activity. According to this physico-mathematical method, it is possible to define amino acids in the sequence which are predicted to be the most critical for protein function. Using sperm whale myoglobin, human hemoglobin and hen egg white lysozyme as model protein examples, sets of predicted amino acids, or so-called 'hot spots', have been identified within the tertiary structure. It was found for each protein that the predicted 'hot spots', which are distributed along the primary sequence, are spatially grouped in a dome-like arrangement over the active site. The identified amino acids did not correspond to the amino acid residues which are involved in the chemical reaction site of these proteins. It is thus proposed that the resonant recognition model helps to identify amino acid residues which are important for the creation of the molecular structure around the catalytic active site and also the associated physical field conditions required for biorecognition, docking of the specific substrate and full biological activity. 相似文献
174.
KCl or LiCl, when added in 100 mM concentrations to cardiac sarcoplasmic reticulum incubated at 17 degrees C with 5 micron [gamma-32P]ATP, 1 mM MgCl2, and 9.1 micron M Ca2+, increased the apparent phosphorylation rate constant from 14.5 s-1 to 23.8 s-1 (100 mM LiCl) or to 44.1 s-1 (100 mM KCl). These same monovalent cations also increased the apparent rate constant for the hydrolysis of the phosphorylated sarcoplasmic reticulum from 0.51 s-1 to 1.12 s-1 (100 mM LiCl) or to 1.71 s-1 (100 mM KCl). Although there was a small burst in Pi production, rate constant of 0.97 s-1, when 100 mM KCl was added, the burst when LiCl or no monovalent cation was added was either nonexistent or so small as to make its detection unreliable. KCl thus appears to induce an intermediate which is either nonexistent when omitted or in such low concentration as not to be readily detected. 相似文献
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Ciayadi R Potdar M Walton KL Harrison CA Kelso GF Harris SJ Hearn MT 《Bioorganic & medicinal chemistry letters》2011,21(18):5642-5645
Novel inhibitors of TGF-β1 and activin A signalling based on a 2-aryl-4-(3-(pyridin-2-yl)-1H-pyrazol-4-yl)pyridine pharmacophore have been synthesised. Compounds containing phenyl or aromatic nitrogen heterocycle substituents inhibited both types of signalling with HEK-293T cells in culture, with a selectivity preference for TGF-β1. Synthetic compounds containing pyridin-3-yl, pyrazol-4-yl, pyrazol-1-yl or 1H-imidazoyl-1-yl substituents exhibited structural and functional attributes suitable for further investigation related to the development of more potent TGF-β inhibitors. 相似文献
177.
Rondine J Allen Evan P Brenner Caitlin E VanOrsdel Jessica J Hobson David J Hearn Matthew R Hemm 《BMC genomics》2014,15(1)
Background
The reliable identification of proteins containing 50 or fewer amino acids is difficult due to the limited information content in short sequences. The 37 amino acid CydX protein in Escherichia coli is a member of the cytochrome bd oxidase complex, an enzyme found throughout Eubacteria. To investigate the extent of CydX conservation and prevalence and evaluate different methods of small protein homologue identification, we surveyed 1095 Eubacteria species for the presence of the small protein.Results
Over 300 homologues were identified, including 80 unannotated genes. The ability of both closely-related and divergent homologues to complement the E. coli ΔcydX mutant supports our identification techniques, and suggests that CydX homologues retain similar function among divergent species. However, sequence analysis of these proteins shows a great degree of variability, with only a few highly-conserved residues. An analysis of the co-variation between CydX homologues and their corresponding cydA and cydB genes shows a close synteny of the small protein with the CydA long Q-loop. Phylogenetic analysis suggests that the cydABX operon has undergone horizontal gene transfer, although the cydX gene likely evolved in a progenitor of the Alpha, Beta, and Gammaproteobacteria. Further investigation of cydAB operons identified two additional conserved hypothetical small proteins: CydY encoded in CydAQlong operons that lack cydX, and CydZ encoded in more than 150 CydAQshort operons.Conclusions
This study provides a systematic analysis of bioinformatics techniques required for the unique challenges present in small protein identification and phylogenetic analyses. These results elucidate the prevalence of CydX throughout the Proteobacteria, provide insight into the selection pressure and sequence requirements for CydX function, and suggest a potential functional interaction between the small protein and the CydA Q-loop, an enigmatic domain of the cytochrome bd oxidase complex. Finally, these results identify other conserved small proteins encoded in cytochrome bd oxidase operons, suggesting that small protein subunits may be a more common component of these enzymes than previously thought.Electronic supplementary material
The online version of this article (doi:10.1186/1471-2164-15-946) contains supplementary material, which is available to authorized users. 相似文献178.
179.
Removal of cleavage slow points from affinity tags used in the IMAC purification of recombinant proteins
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Jane T. Mooney Dale Fredericks Thorkild Christensen Milton T. W. Hearn 《Biotechnology journal》2014,9(8):1023-1032
The complete enzymatic removal of affinity tags from tagged recombinant proteins is often required but can be challenging when slow points for cleavage exist. This study documents a general approach to remove N‐terminal tags from recombinant proteins specifically designed to be efficiently captured by IMAC resins. In particular, site‐directed mutagenesis procedures have been used to modify the amino acid sequence of metal binding tags useful in IMAC purifications of recombinant proteins with the objective to increase cleavage efficiency with the exopeptidase, dipeptidyl aminopeptidase 1. These tags were specifically developed for application with borderline metal ions, such as Ni2+ or Cu2+ ions, chelated to the immobilized ligands, 1,4,7‐triazacyclononane (tacn) and its analogs. Due to the ability to control cleavage site structure and accessibility via site directed mutagenesis methods, these procedures offer considerable scope to obtain recombinant proteins with authentic native N‐termini, thus avoiding any impact on structural stability, humoral and cellular immune responses, or other biological functions. Collectively, these IMAC‐based methods provide a practical alternative to other procedures for the purification of recombinant proteins with tag removal. Overall, this approach is essentially operating as an integrated down‐stream purification capability. 相似文献
180.
Jonathan Hearn 《American anthropologist》1999,101(1):216-217
Europe in the Anthropological Imagination. Susan Parman. ed. Saddle River, NJ: Prentice Hall, 1998.274 pp. 相似文献