52 results about "Aminoacylation" patented technology

The drug action mechanisms and core techniques of the present invention are summarized in figure 1. Specifically, the invention provides malignant denatured proteins, such as mutant Huntingtin proteinor alpha-synuclein, stick together to grow into oligomer aggregates (1, 2), fibrillar aggregates (3), and ultimately inclusion bodies (4). Young neuronal cells produce a large quantity of Nt-Arg through N-terminal arginylation (5) of endoplasmic reticulum chaperones, such as BiP, and thereafter, arginylated BiP (R-BiP) comes into the cytoplasm and binds with denatured proteins (6). Nt-Arg of R-BiP, as a ligand, binds with the ZZ domain of p62 (7) to induce the structural activation of p62 (8) while the ordinarily closed inactive form of p62 is changed with an open form thereof, and thus PB1 and LC3-binding domains are exposed. On the basis of oligomerization (9) by the PB1 domain, p62 binds with the denatured protein aggregates to be concentrated to autophagically degradable aggregates, that is, p62 bodies (10). Thereafter, p62 completes autophagy targeting (11) and lysosomal proteolysis through binding with LC3 protruding on the autophagosomal membranes. In young neuronal cells, theautophagic proteolysis occurring through steps 5-11 is strong, and thus the cytotoxic protein aggregates (1-5) do not accumulate, but in aged neuronal cells, the autophagic proteolysis occurring through steps 5-11 is weakened, and thus the protein aggregates (1-5) accumulate, resulting in a vicious cycle. The present invention attempts to effectively remove Huntingtin and alpha-synuclein protein aggregates and the like by artificially activating p62 using low-mass ligands of the p62 ZZ domain (12, 13). Specifically, p62 binding the ligands through step 12 promotes p62-R-BiP-denatured protein oligomerization (9) and autophagy aggregate formation (10). In addition, the ligand-62 conjugates step 13 act as autophagy activators (14), to promote LC3 synthesis, the conversion of LC3-I into LC3-II, and the like, thereby promoting the formation of autophagosomes (15).

An aminoacylation / translation (AIT) system based on the protein synthesis system from the pathogen Pseudomonas aeruginosa, was used to screen chemical compounds for identifying inhibitors of protein synthesis. This system includes elongation factors: EF-Tu, EF-Ts and EF-G, aminoacyl tRNA synthetase (aaRS) specific for phenylalanine, PheRS, and ribosomes isolated from cultures of Pseudomonas aeruginosa. Compounds identified using this assay have been shown to contain broad spectrum activity against both Gram+ and Gram− pathogens. Methods of using the identified compounds, as well as derivatives and analogues of these compounds, as antimicrobial agents against bacterial infections are described.

The invention relates to an aminoacyl-tRNA synthetase mutant which is orthogonal aminoacyl-tRNA synthetase. The amino acid sequence of the synthetase is selected from a group composed of an amino acid sequence presented in SEQ ID NO:4 and conservative variants of the amino acid sequence presented in SEQ ID NO:4, and the conservative variants have enzyme activity the same as the amino acid sequence presented in SEQ ID NO:4. The invention further provides a 3,5-difluoro-tyrosine translation system which comprises (i) 3,5-difluoro-tyrosine, (ii) the orthogonal aminoacyl-tRAN synthetase, (iii) orthogonal tRNA and (iv) nucleic acid of encoded target protein, wherein the orthogonal aminoacyl-tRAN synthetase carries out prior aminoacylation on the orthogonal tRNA through the 3,5-difluoro-tyrosine, and the nucleic acid contains at least one selective codon for specific recognition of the orthogonal tRNA.

It is intended to provide a method of chemically synthesizing an aminoacyl tRNA which is completely different from the existing methods, namely, a highly efficient and practically usable method for synthesizing an aminoacyl tRNA whereby any nonnatural amino acid can be conveniently aminoacylated without a need for genetic engineering techniques and detected without resort to a radioactive isotope. The above-described aminoacylation method comprises enclosing a tRNA and an amino acid in the vicinity of the micelle-water interface and bringing them close to each other to thereby react the same, or providing between them a peptide nucleic acid specifically binding to the tRNA as an antisense molecule and bringing them close to each other to thereby react the same.