11 results about "Substrate recognition" patented technology

The invention belongs to the technical field of organic synthetic chemistry, and discloses a method for efficiently synthesizing methyl 2-chloro-3-oxovalerate. According to the method, high-selectivity chlorination is performed on a beta-site C-H bond of methyl 3-oxovalerate under a mild condition by taking sodium chloride as a chlorine source and adopting a dual-catalysis system consisting of engineered halogenase and a visible light-sensitive catalyst under visible light irradiation. The engineered halogenase is a HalB triple mutant, and the affinity and regioselectivity of a substrate are remarkably improved; the photosensitive catalyst is a ruthenium bipyridine complex and synergistically drives generation of chlorine free radicals. According to the invention, a brand new biological-photochemical synergistic catalysis platform is constructed by organically combining the substrate recognition capability of the engineered halogenase and an electron transfer mechanism driven by visible light catalysis, so that the contradiction among selectivity, mildness and atomic efficiency of a traditional chlorination method is solved; and the method can be expanded to precise synthesis of other beta-functionalized ketone ester compounds.

The invention relates to the cross technical field of artificial intelligence and bioinformatics, in particular to a multi-modal enzyme kinetic parameter prediction method of an adaptive protein language model. Aiming at the problems that an existing processing method is lack of enzymatic reaction dynamic mechanism modeling and insufficient in three-dimensional structure utilization, the invention provides the following technical scheme: step 1, acquiring and preprocessing multi-modal data; 2, protein language model and molecular fingerprint feature extraction; 3, carrying out substrate recognition feature fusion based on cross attention; step 4, extracting conformation adaptive features based on the hybrid expert network; 5, correcting enzyme-substrate distribution alignment characteristics; step 6, kinetic parameter regression prediction; step 7, constructing a multi-objective loss function; and step 8, model training and parameter optimization. Through an enzyme reaction bridging adapter and an enzyme-substrate distribution alignment technology, high-precision prediction of enzyme kinetic parameters is realized, and cross-data-set accuracy and robustness are remarkably improved.

The invention provides a nucleic acid in-situ dual-channel fluorescence detection probe group and system based on cleavage type DNAzyme and application of the nucleic acid in-situ dual-channel fluorescence detection probe group and system. The probe group comprises at least two probes, each probe comprises a recognition activation region and a signal generation region, the recognition activation region sequentially comprises a target nucleic acid recognition arm, a catalytic core sequence and a substrate recognition arm, and each probe can be paired with at least another probe; a substrate recognition arm and a catalytic core sequence on the paired probe are two cleavage type DNAzyme subunits formed by DNAzyme, and the DNAzyme with catalytic activity is formed by self-assembly in the presence of target nucleic acid; the signal generation region comprises at least one substrate of DNAzyme, two ends of the substrate are respectively modified with a fluorophore and a quenching group, the substrate is complementary with a substrate recognition arm, and the fluorophore modified on the substrate of each probe is different. According to the invention, in-situ real-time dynamic imaging, high-specificity and high-sensitivity detection and targeted regulation of target nucleic acid are realized.

DCAF15 is a substrate recognition (adaptor) protein of E3 ubiquitin ligase. Disclosed herein are compounds that recruit ubiquitin ligase CRL4DCAF15, to a target RNA recognition motif (RRM), causing its degradation. Also disclosed herein are compositions and methods of use in treating associated disorders and diseases.

The invention discloses a molecular docking-based glycosyl transferase substrate catalytic specificity multi-dimensional evaluation method, system, equipment and medium, and belongs to the technical field of protein structure simulation evaluation, and the method comprises the following steps: determining glycosyl transferase, and predicting a corresponding protein three-dimensional structure; performing molecular docking simulation based on the predicted protein three-dimensional structure; the catalytic specificity of different substrates is evaluated according to the difference between receptor substrate channels and ligands; and displaying a docking result through visual software. According to the method, a set of complete standardized operation program is formed, a clear methodology guide is provided for similar research, on the basis of traditional docking scoring, three-dimensional space size measurement analysis of the acceptor and the ligand is creatively used as a key screening index, multi-dimensional evaluation from the bonding strength to the space suitability is achieved, and the evaluation accuracy is improved. A new visual angle is provided for understanding an enzyme substrate recognition mechanism.

The application provides a preparation method of an engineered DEAR nucleic acid manipulation system, and the preparation method comprises at least one selected from the following: lengthening a substrate recognition region in a RNA molecule of an original DEAR nucleic acid manipulation system to have a length of 7-14 nucleotides; adding a recruitment sequence to a 3' end of a RNA molecule of the original DEAR nucleic acid manipulation system; respectively arranging a first dimerization motif and a second dimerization motif in a III domain in a first RNA molecule and a second RNA molecule from the original DEAR nucleic acid manipulation system, so that the first RNA molecule and the second RNA molecule form a heterodimer, and the like. The preparation method of the engineered DEAR nucleic acid manipulation system provided by the application further improves the specificity and cleavage activity of the DEAR nucleic acid manipulation system based on an RNA ribozyme.

The invention relates to the technical field of biology, in particular to an LbCas12a variant and application thereof in nucleic acid detection. According to the invention, a series of LbCas12a mutants with differentiated enzyme digestion activities are obtained by modifying key amino acid sites related to substrate recognition and conformation activation in LbCas12a protein. Compared with an existing method, a one-tube nucleic acid detection system based on the mutant has better system stability, sensitivity, specificity and universality, and high-sensitivity one-tube nucleic acid detection can be achieved under the condition that exogenous addition is not needed, and special consumables or complex equipment are not needed. The mutant set provides a more advantageous tool for the on-site application of a CRISPR (clustered regularly interspaced short palindromic repeats) detection system.

This invention discloses an RNA ribozyme-based DEAR nucleic acid manipulation system and its applications. The invention provides a DEAR nucleic acid manipulation system comprising a bacterial-derived C-type group II intron RNA molecule, the RNA molecule comprising a substrate recognition region that hybridizes with a target sequence in a target nucleic acid. The RNA ribozyme-based DEAR nucleic acid manipulation system provided in this invention avoids problems in the CRISPR-Cas system, such as reduced transfection efficiency due to protein molecule oversize and potential immunogenicity caused by the Cas protein. The RNA ribozyme-based DEAR nucleic acid manipulation system provided in this invention is capable of cleaving DNA and RNA.

The present application relates to the field of tumor immunotherapy, discloses a kind of ribozyme for targeted inhibition of PD-L1 gene expression and its preparation method and application, specifically relates to a kind of engineered Pistol ribozyme and its use in targeted inhibition of PD-L1 (CD274) gene expression.The ribozyme is RNA molecule connected by ribonucleotide monomer, contains the substrate recognition region of the conserved structural framework maintaining catalytic activity and PD-L1 transcript complement;Conserved skeleton covers P1, P2, P3 region and false node area, and contains catalytic related conserved base site, substrate recognition region can be designed as required, can recognize and cut the targeted site containing GU dinucleotide in PD-L1 transcript, reduce the PD-L1 expression of tumor cell in post-transcriptional level.The PD-L1 transcript includes mature mRNA, precursor mRNA, intron retention transcript and variable splicing transcript.Different from existing immune checkpoint blockade therapy, the present application can sustainably inhibit endogenous PD-L1 in tumor cell, relieve immune suppression, enhance anti-tumor immune response, and also provide the preparation method of the ribozyme and its pharmaceutical composition, and application prospect is good.

The present invention relates to the modification of effector proteins (e.g., E3 ligase) to alter their activity (e.g., induced functional obtainment). The present invention relates to methods of modifying effector proteins by inserting peptide motifs or libraries of peptide motifs into effector proteins, libraries and methods of screening with modified effector proteins to identify new protein functions, i.e., new protein-protein interactions. For example, the present invention relates to a method of identifying a modified effector protein (e.g., an E3 ligase) having an induced change in substrate recognition activity, identified by measuring a change in substrate binding activity and / or degradation of a target protein. For example, the present invention enables the identification of new E3 ligases: Protein Of Interest (POI) interfaces, which can be used in drug discovery and development, such as monovalent degrading agent drugs such as molecular glue.