86 results about "Protein stability" patented technology

Provided herein are technologies for framing and evaluating biological sequence-based analysis tasks in a unified, natural-language-based, text in and text out format. Among other things, methods and systems of the present disclosure provide machine-learning technologies for combining biological sequence data, representing, for example, DNA, RNA, and protein sequences, with natural language, conversational style prompts that set out particular analysis tasks to be performed on the biological sequence data. This approach, for example, allows complex analysis tasks, including, but not limited to, identification of various sequence modifications, genes, and regulatory elements in DNA sequences, and quantification of properties such as degradation propensity of RNA and protein stability, to be input to a machine learning model in a uniform text-based format and for output to be generated in a same, unified, text-based format.

The present disclosure provides drug responsive domains derived from human carbonic anhydrase 2 that can modulate protein stability for human interleukin 15 (IL15) payloads, as well as compositions and methods of use thereof.

The invention provides a protein mass spectrum data analysis method and system based on protein stability. The method comprises the following steps: determining scale constraint parameters of a sliding window during smoothing of protein mass spectrum data according to local noise variances at data points in the protein mass spectrum data; carrying out loss constraint on the edge of a protein stability characteristic peak in the protein mass spectrum data based on the scale constraint parameter in combination with a preset sliding window to obtain edge retention data of the protein stability characteristic peak; determining a fuzzy membership degree of each data point belonging to a protein stability characteristic peak according to a plurality of pre-identification peaks in the edge retention data and spatial distribution characteristics of each data point in the edge retention data; and performing baseline correction on the edge retention data based on Bayesian probability in combination with all fuzzy membership degrees, and further extracting peptide fragment signal peaks of the protein. According to the technical scheme provided by the invention, the effective peptide fragment signal peak in the protein mass spectrum data can be analyzed in a non-stationary background noise state.

A coronavirus-targeting antibody nasal spray comprising: a coronavirus-targeting antibody or antigen-binding fragment, a buffer, a stabilizer, a surfactant, and a thickening agent, and optionally further comprising a preservative. The nasal spray can improve the physicochemical stability of the coronavirus-targeting antibody, and the nasal spray can maintain protein stability under conditions such as room temperature, long-term refrigeration, and repeated freeze-thawing, thereby improving the safety and effectiveness of clinical use.

The present disclosure relates to the use of a maltose dependent degron to control stability of a protein of interest fused thereto at the post-translational level. The present disclosure also relates to the use of a maltose dependent degron in combination with a maltose-responsive promoter to control gene expression at the transcriptional level and to control protein stability at the post-translational level. The present disclosure also relates to the use of a stabilization construct that couples expression of a cell-growth-affecting protein with the production of non-catabolic compounds. The present disclosure further relates to the use of a synthetic maltose-responsive promoter. The present disclosure further provides compositions and methods for using a maltose dependent degron, a maltose-responsive promoter, and a stabilization construct, either alone or in various combinations, for the production of non-catabolic compounds in genetically modified host cells.

The invention relates to application of a GSTP1 inhibitor in preparation of a medicine for treating chronic myelogenous leukemia, and belongs to the technical field of biomedicine. According to the application, it is found for the first time that interaction exists between CML core driving proteins BCR-ABL1 and GSTP1, GSTP1-mediated BCR-ABL1 protein glutathione modification can be effectively inhibited through targeted inhibition of the expression level of the GSTP1, the stability of the BCR-ABL1 protein is reduced, CML cell proliferation is inhibited, and CML cell apoptosis is promoted, so that the effect of treating the chronic myeloid leukemia is achieved. Therefore, the GSTP1 can be used as a treatment target, the GSTP1 inhibitor Ezatiostat is used for preparing the medicine for treating the chronic myelogenous leukemia, a theoretical basis and possibility are provided for developing a GSTP1 inhibitor and TKI combined medication scheme in the future, synergistic interaction is expected, and prognosis of a patient is further improved.

Disclosed herein are peptide hydrogel-based formulations. In some embodiments, the peptide hydrogel-based formulations encapsulate antibodies. In some embodiments, the peptide hydrogel-based formulations prevent formation of high molecular weight protein species.

The invention belongs to the technical field of action target screening and identification, and particularly relates to a method for screening and identifying a Nav channel protein binding agent based on ligand-induced protein stability change. The method comprises the following steps: firstly, carrying out mild lysis on cells of high-expression voltage-gated sodium ion channel protein (Nav channel protein) to obtain lysate containing active Nav channel protein, carrying out contrast incubation on the lysate, a compound to be detected and a solvent, carrying out restrictive enzymolysis by using protease in the presence of a surfactant, and carrying out freeze-drying to obtain the high-expression voltage-gated sodium ion channel protein (Nav channel protein). Representing the retention amount of the target protein and the internal reference protein, and evaluating the protective capability of the compound to be detected on degradation of the target protein to judge the affinity. The method has the advantages of high identification speed, economy and high efficiency, overcomes the problems that the existing Nav channel protein binding agent screening method is tedious, high in technical difficulty and depends on special large instruments, and can be used for screening action targets without specially constructing cells and purifying proteins by large instruments, so that the screening efficiency is effectively improved, and the screening cost is effectively reduced.

This invention discloses a mutant, gene, and recombinant bacteria of a highly thermostable single-chain sweet protein. The mutant is characterized by the following amino acid sequences shown in SEQ ID NO.1: isoleucine (I) at position 5 is mutated to glutamic acid (E), glutamic acid (E) at position 23 is mutated to alanine (A), isoleucine (I) at position 26 is mutated to arginine (R), cysteine ​​(C) at position 41 is mutated to alanine (A), tyrosine (Y) at position 65 is mutated to isoleucine (I), glycine (G) at position 83 is mutated to arginine (R), and asparagine (N) at position 90 is mutated to glutamic acid (E). Based on structural analysis and computationally assisted design for protein stability, this invention achieves site-directed mutagenesis of the single-chain sweet protein, resulting in a highly thermostable mutant. Compared to the original single-chain sweet protein, the thermostable mutant of this invention exhibits higher T... m It was heated by more than 20.4°C while maintaining its sweetness.

Fusion proteins and methods for conditionally fine-tuning transposase protein stability and activity for clinical applications are provided herein

The application discloses a methionine adenosyltransferase mutant and application of the mutant in immobilized reaction, the mutant is obtained by mutating an amino acid sequence of wild-type Escherichia coli methionine adenosyltransferase, the mutant enhances the interaction force between subunits by introducing a disulfide bond at a specific position; and / or, the protein stability is enhanced by constructing a connecting peptide between adjacent subunits. The mutant provided in the application enhances the thermal stability and catalytic efficiency of the enzyme, reduces the optimum reaction pH value, reduces the generation of by-products and the precipitation of a reaction solution. The mutant enzyme can maintain high activity at a temperature of about 40 DEG C, and is particularly suitable for industrial production. In addition, the immobilized form of the mutant enzyme exhibits good stability and reusability in continuous use, significantly reduces production cost, simultaneously reduces environmental pollution, improves the yield of SAM, is more suitable for large-scale industrial production, and has more market competitiveness.

The application discloses a mutant and gene of single-chain sweet protein with high thermal stability, wherein the mutant is obtained by mutating isoleucine (I) at the 5th position, glutamic acid (E) at the 23rd position, isoleucine (I) at the 26th position, tyrosine (Y) at the 65th position, glycine (G) at the 83rd position and asparagine (N) at the 90th position in the amino acid sequence shown in SEQ ID NO. 1 to alanine (A), arginine (R), isoleucine (I), glutamic acid (E) and aspartic acid (D) respectively; the application is based on structural analysis and protein stability calculation aided design, and the single-chain sweet protein is subjected to site-directed mutagenesis, so that the mutant of single-chain sweet protein with high thermal stability is obtained; compared with the single-chain sweet protein, the mutant of single-chain sweet protein with high thermal stability has a T m increase of 18.4 DEG C or more and maintains sweetness.

The invention provides application of an expression inhibitor of KMT5A in preparation of a medicine for preventing or treating esophageal squamous carcinoma. In-vitro experiments prove that ICA can inhibit proliferation of esophageal squamous carcinoma cells in a concentration-dependent manner and remarkably promote cell apoptosis. The action mechanism of the ICA is closely related to the direct targeting KMT5A protein, and experiments show that the ICA can reduce the expression level of the KMT5A and influence the protein stability of the KMT5A. Clinical sample analysis further shows that KMT5A is highly expressed in esophageal cancer tissues, and ICA treatment in animal experiments significantly reduces the tumor volume. The invention provides a new targeted treatment strategy for esophageal squamous carcinoma, and has important clinical transformation value.

Embodiments of the present application provide a protein stability prediction method and device, computer equipment and a storage medium. The method comprises: obtaining a first to-be-tested sequence corresponding to an original protein and a second to-be-tested sequence corresponding to a mutant protein; determining a plurality of first target residue features from the first to-be-tested sequence according to the attention mechanism of the protein language model and extracting the global features thereof to obtain first virtual structure microenvironment features; similarly, obtaining second virtual structure microenvironment features corresponding to a plurality of second target residue features; determining a first difference based on the first virtual structure microenvironment features and the second virtual structure microenvironment features, and determining a second difference based on a first global sequence feature corresponding to the first to-be-tested sequence and a second global sequence feature corresponding to the second to-be-tested sequence; and predicting based on the first difference and the second difference to obtain a protein stability prediction result. In this way, the accuracy of predicting the stability of the protein can be improved.

The invention relates to the technical field of gene engineering and enzyme engineering, and discloses a sugarcane sucrose phosphate synthase truncation and application thereof, and the amino acid sequence of the truncation is as shown in SEQ ID NO.4. The sugarcane sucrose phosphate synthase truncation provided by the invention is obtained by cutting off 37th-42nd amino acids on the basis of a wild type sugarcane sucrose synthase (ScSPSB) gene from sugarcane cinnabar 42, and compared with the wild type ScSPSB gene, the sugarcane sucrose phosphate synthase truncation has higher catalytic activity and stability. According to the invention, a wild type ScSPSB gene is subjected to truncation transformation to obtain the sugarcane sucrose phosphate synthase truncation with higher catalytic activity and higher protein stability, so that a foundation is laid for studying transcription and expression mechanisms of sugarcane sucrose phosphate synthase and further discussing an accumulation mechanism of sucrose; and a theoretical and practical basis is provided for researching the biological function of the sucrose phosphate synthase and improving crop varieties by applying the sugarcane gene.

The invention provides a method for efficiently preparing high-purity NudCD3 protein in an in-vitro eukaryotic cell. The method comprises the following steps: synthesizing a target gene; carrying out enzyme digestion on the target gene and the pLEXmMBP vector, and connecting to obtain a recombinant vector; transforming the recombinant vector into competent cells, culturing, and extracting plasmids; transfecting an HEK 293F cell, culturing, and collecting the cell; resuspending the cells in a lysis buffer solution, carrying out ultrasonication and centrifugation, and taking a supernatant; and carrying out affinity chromatography column purification, Prescission protease digestion and molecular sieve purification to finally obtain the NudCD3 protein with the purity of more than 90%. The problems that in the prior art, NudCD3 protein is difficult to express in quantity and insufficient in purity are solved, high-purity protein is provided for structural biology research, drug screening and antibody preparation, and the method has the advantages of being easy to operate, high in yield, high in protein stability and the like.

The invention relates to the technical field of protease mutation, and discloses a bisabolol synthase mutant with high catalytic activity and application of the bisabolol synthase mutant. According to the method, alpha-bisabolol synthase derived from compositae herbaceous plant globe artichoke is taken as wild enzyme, mutants are virtually screened by applying a protein language model SaProt, a deep learning model Catapro and a protein stability prediction tool Dynaamt, and the effect of producing bisabolol is verified through experiments; according to the present invention, the mutants obtained by mutation of 11 specific amino acid sites such as D125N, F159Y, F159K, N346D, T372S, Y432F, N346S, D184E, N157E, E374A and T258S are obtained through the mutation of the above 11 specific amino acid sites; wherein through 96-hour shake flask fermentation and gas chromatography detection, the bisabolol yields of the obtained single-point mutation T258S and double-point mutation T258S / F159K are respectively increased by 72.6% and 90.2% compared with the wild type yield; the Kcat / Km value of the two-point mutation T258S / F159K is 4.57 times that of a wild type.

The application discloses a mutant of single-chain sweet protein with high thermal stability and a gene of the mutant, wherein the mutant is obtained by mutating isoleucine (I) at the 5th position, glutamic acid (E) at the 23rd position, isoleucine (I) at the 26th position, tyrosine (Y) at the 65th position, glycine (G) at the 83rd position and asparagine (N) at the 90th position in the amino acid sequence shown in SEQ ID NO. 1 to alanine (A), arginine (R) and aspartic acid (D) respectively. m The Tm of the mutant of single-chain sweet protein with high thermal stability is increased by more than 19.4 DEG C, and the sweetness is maintained.

The invention provides an application of an HIF-3alpha-3 gene or a protein coded by the HIF-3alpha-3 gene in preparation of a medicine for preventing and treating bladder cancer, and belongs to the technical field of biological medicines. The invention reveals that HIF-3alpha-3 in bladder cancer up-regulates PHD3 gene expression from a transcriptional regulation level, so that the hydroxylation level of HIF-1alpha in bladder cancer cells is increased, and the protein stability of HIF-1alpha is reduced for the first time. Meanwhile, overexpression of the HIF-3alpha-3 gene or the coded protein can effectively inhibit proliferation and in-vivo tumorigenicity of bladder cancer cells. Therefore, the invention provides the application of the HIF-3alpha-3 gene or the protein coded by the HIF-3alpha-3 gene or the PHD3 gene or the protein coded by the PHD3 gene in preparation of medicines for preventing and treating bladder cancer.

This invention belongs to the field of plant genetic engineering and relates to the application and method of reducing the expression of the Zm7649 gene in improving plant stress resistance. This invention utilizes an EMS mutant with premature termination of expression. Zm7649-ems Heterologous overexpression with Arabidopsis thaliana Zm7649-OE Verified Zm7649 The function of negatively regulating salt stress and high temperature stress was studied, and its preliminary analysis was performed. Zm00001d027649 The molecular mechanism regulating stress involves the interaction between Zm7649 and a maize catalase (Zm4848), with ubiquitination degrading Zm4848 and reducing its protein stability. This invention provides new genetic resources and theoretical basis for maize stress-resistance breeding or germplasm innovation.

Disclosed are a strain capable of expressing an exogenous protein, a recombinant human-derived collagen, a synthesis method, and the use. The present invention first provides the strain capable of expressing the exogenous protein, and the strain is proline-auxotrophic Escherichia coli and can efficiently express the exogenous protein. Using the strain as a host to express the recombinant human-derived collagen with supplementation of exogenous proline and hydroxyproline can achieve accurate regulation and control on the collagen hydroxylation rate by means of regulating the ratio of proline to hydroxyproline in a culture medium. By means of the synthesis method, a recombinant collagen of which the hydroxylation rate is approximate to that of natural human collagen can be obtained and has better cell adhesion and protein stability.

Provided herein are technologies for framing and evaluating biological sequence-based analysis tasks in a unified, natural-language-based, text in and text out format. Among other things, methods and systems of the present disclosure provide machine-learning technologies for combining biological sequence data, representing, for example, DNA, RNA, and protein sequences, with natural language, conversational style prompts that set out particular analysis tasks to be performed on the biological sequence data. This approach, for example, allows complex analysis tasks, including, but not limited to, identification of various sequence modifications, genes, and regulatory elements in DNA sequences, and quantification of properties such as degradation propensity of RNA and protein stability, to be input to a machine learning model in a uniform text-based format and for output to be generated in a same, unified, text-based format.

The present application relates to the construction method of the ophthalmic protein drug sustained-release preparation, the ophthalmic protein drug is contacted with the weakly interacting excipient of pharmaceutically acceptable, and the protein pre-holding complex is formed;The first hydration excipient and the second hydration excipient are sequentially added to the protein pre-holding complex, and the layered hydrated protein complex is formed;The layered hydrated protein complex is dispersed in the first continuous phase of the ionizable group-containing pharmaceutically high polymer, and the ion pairing release blocking layer is formed in the periphery by adjusting the ion environment, and the intermediate is obtained;The intermediate is dispersed in the second continuous phase of the ophthalmic pharmaceutically acceptable, and the ophthalmic protein drug sustained-release preparation is prepared.The ophthalmic protein drug is pre-held with the weakly interacting excipient, the differentiated occupation area and the layered hydration structure are constructed, the protein stability is improved;Through the stage ion regulation, the pharmaceutically high polymer forms the non-closed ion pairing release blocking layer, the protein release fine control is realized, the drug efficacy is prolonged and the curative effect is improved.

The invention discloses a mutant of HPV53 E6 protein and application of the mutant. Compared with wild type E6, the mutant has the following three mutation sites: K79W, S133P and N8K, the K79W mutation design enhances the stability of a hydrophobic space and forms new hydrophobic interaction with Y108 / H130, S133P enables the flexibility of a loop region to be more stable, N8K mutation forms a salt bridge with E10 / E11, and the relation between the loop region and helix is enhanced. The mutation designs can maintain the stability of the whole structure, significantly improve the protein stability and immunogenicity, break through the technical bottlenecks of low in-vivo expression quantity and weak immunogenicity of the natural E6 protein, and provide a new way for immunotherapy of HPV53 related lesions.

The application discloses a hair loss prevention shampoo targeting CD133 positive hair follicle dermal papilla cells and a preparation method thereof. The shampoo comprises a hair follicle targeting single-domain antibody-active peptide fusion, a liposome-encapsulated compound plant extract, a biological surfactant, a protein stability surfactant and an auxiliary material. The fusion is formed by fusing an anti-CD133 VHH single-domain antibody and a 5alpha-reductase inhibiting peptide DK-HP-7 through a flexible connecting peptide. Based on the discovery that CD133 positive dermal papilla cells are a key cell group driving hair follicle formation, the application uses single-domain antibodies to achieve precise targeting of active ingredients. The DK-HP-7 designed through molecular docking has an IC 50 of 4.28 μM. The rhamnolipid and Kolliphor HS 15 complex system can make the fusion protein activity retention rate higher than 89% after being stored at 40 ℃ for 3 months. The shampoo can significantly antagonize dihydrotestosterone-induced hair follicle growth inhibition, promote hair follicles to enter the growth phase and increase hair follicle density, and has good skin safety and stability.

This application provides a protein stability detection probe based on ascorbate peroxidase 2 and its application. The detection probe includes ascorbate peroxidase 2 displayed on the cell surface and an anchoring protein that anchors ascorbate peroxidase 2 to the cell. Ascorbate peroxidase 2 has an insertion site for the target protein, into which the target protein inserts. Ascorbate peroxidase 2 is divided into N-terminal and C-terminal ascorbate peroxidase 2, and the target protein is linked to both ends of the C-terminal ascorbate peroxidase 2 via flexible linkers. The intensity of the fluorescence signal generated by the cascade reaction catalyzed by the probe in this application has a good linear relationship with the stability of the inserted test protein mutant, enabling high-throughput identification of protein mutant stability and showing promising application prospects.

An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.

A recombinant TEV protease, a preparation method therefor, and use thereof, relating to the technical field of biology. Specifically provided are four recombinant TEV proteases derived from a cysteine protease of tobacco etch virus. In each of the four recombinant TEV proteases, the N-terminus is fused with an 8His-CL7-GGS tag, and the C-terminus is fused with a polyarginine tag. Compared to a wild-type TEV protease, the provided four recombinant TEV proteases not only can overcome defects such as self-cleavage, poor solubility, and low activity during expression and purification of the wild-type TEV protease, but also have improved protein yield 5-6 times higher than that of the wild-type TEV enzyme, and enzyme activity 10-20 times higher than that of the wild-type TEV protease. In addition, the engineered TEV protease mutants also possess higher thermal stability with a Tm value 10-30 °C higher than that of the wild-type TEV protease, exhibit better protein stability, have broader application conditions, and are more suitable for large-scale production and industrial use.

The invention relates to an efficient multifunctional protein modification design platform system and an operation method thereof. The efficient multifunctional protein modification design platform system comprises a work command station, a multi-user terminal, a platform website and a function module. The functional module comprises a structure prediction module, a molecular docking module, a molecular dynamics simulation module, a de novo design module, a directed evolution module, an evolution and conservative analysis module and a protein stability analysis module. A user uses a functional module of the platform system to carry out integrated calculation and analysis through a link established by the work command station for the user, and the platform system is used for transformation design of target proteins, including protein-protein complexes, protein-polypeptide combinations, protein-small molecule combinations, functional proteins or enzymes and the like. According to the platform system, a graphical interface easy to operate is designed, operation automation of data processing and operation and protein design data results with high accuracy and success rate are achieved, the platform system is used for improving protein performance, and protein with novel functions is designed and obtained.

Cancer metastasis is the leading cause of cancer related death and involves the spreading of the tumor to distant sites throughout the body. However, there are limited therapies which can effectively target this deadly process. One protein which contributes to cancer metastasis is termed flotillin-1. Its expression is associated with metastasis in several solid tumors. It is known that the addition of a fatty acid can alter this protein's biology and stabilize it (“palmitoylation”). It is herein shown that altering this process can lead to a decrease in the stability of the protein. Thus, targeting the addition of this fatty acid can lead to its degradation and be a way to successfully target this metastasis-inducing protein therapeutically.