54 results about "Chaperone (protein)" patented technology

The invention relates to a fusion protein, an immunogenic composition and a recombinant degenerate vaccine for preventing and treating infection of various pathogenic streptococci, as well as a molecular architecture design, application and the like. According to the invention, three immune antigens, namely an elongation factor Tu (Tuf), a molecular chaperone DnaK and an elongation factor G (fusA), are screened, and it is proved that fusion protein molecules of the three antigens can significantly inhibit tissue lesions caused by infection of different serotypes and different types of streptococci, have good immunogenicity, play roles in immune protection and effective prevention and treatment, and have good application prospects. The bacillus subtilis has the characteristics of broad-spectrum and high-efficiency prevention of streptococcus bacterial infection, and has a wide application prospect.

The invention discloses a chicken infectious anemia subunit vaccine composition as well as a preparation method and application thereof. The vaccine composition contains a mixture of CIAV (Chicken Infectious Anemia Virus) VP1 and VP2 proteins, the mixture of the VP1 and VP2 proteins is prepared by performing in-vitro renaturation on VP1 inclusion body proteins by using a gradient dialysis method, adding pre-purified VP2 proteins in the dialysis process to promote the recovery of key conformation epitopes of the VP1 proteins, and then gradually removing a denaturing agent to obtain the VP1 and VP2 protein mixture. And refolding the VP1 protein to obtain the protein. The soluble VP2 protein is used as a molecular chaperone to recover the VP1 protein from the inclusion body. According to the VP2 protein assisted synergistic refolding strategy, the key conformation epitope of the VP1 protein is recovered by using the natural function of the VP2 protein. Subsequently, the immunogenicity and protective efficacy of the refolded VP1 and VP2 protein mixture are evaluated, and an economic and effective technical means is provided for preventing and controlling chicken infectious anemia.

The invention discloses a recombinant 2.1 d subtype swine fever E2 protein, a subunit vaccine and application of the subunit vaccine, and belongs to the technical field of genetic engineering. According to the 2.1 d subtype swine fever E2 protein, 344-375 amino acids of the 2.1 d subtype swine fever E2 protein are removed, and 1-343 amino acids of the 2.1 d subtype swine fever E2 protein are reserved, so that the most important antigenic epitope with a protective effect of the 2.1 d subtype swine fever E2 protein can be reserved, the space structure of the recombinant 2.1 d subtype swine fever E2 protein is basically not influenced, and the immunogenicity of the 2.1 d subtype swine fever E2 protein can be reserved to the greatest extent; in addition, the recombinant 2.1 d subtype swine fever E2 protein can be efficiently expressed in a prokaryotic expression system; furthermore, the soluble expression level of the recombinant 2.1 d subtype swine fever E2 protein can be remarkably improved through co-expression of the molecular chaperone, and after the recombinant 2.1 d subtype swine fever E2 protein is prepared into a subunit vaccine, the subunit vaccine has a good protection effect on 2.1 d subtype swine fever virus infection.

The invention provides a method for improving the expression level of a meat toxin light chain. The method comprises the following steps: connecting a dissolution-promoting tag to an N end or a C end of meat toxin light chain protein to form soluble meat toxin light chain recombinant protein; the dissolution-promoting tag is a wild dissolution-promoting tag NT11 or a mutant protein mut7 of the wild dissolution-promoting tag NT11, enzyme digestion removal is not needed, the activity of a meat toxin light chain is hardly influenced and interfered, and the fusion protein can keep 93% of substrate enzyme digestion activity. The method further comprises overexpressing a molecular chaperone htpG in the genome of the host bacterium. According to the invention, a BoNT / A-LC high-efficiency expression system is constructed through multi-strategy optimization, so that the yield of BoNT / A-LC is increased by nearly 20% compared with that of wild type NT11-Lc, the yield reaches 647 mg / L through overexpression of a molecular chaperone htpG, and after combinatorial optimization, the yield is 8.5 times that of only expressed BoNT / A Lc, 1.79 times that of NT11-Lc and 1.43 times that of Mut7-Lc.

Disclosed is a novel class of fusion proteins to recruit the cell's innate chaperone machinery, specifically the Hsp70-mediated system, to specifically reduce TDP-43-mediated protein aggregation and associated protein conformational diseases.

In one aspect, the disclosure relates to compounds methods for treating or preventing diseases associated with aberrant condensation of a biomolecule in a subject, the method including at least the step of contacting one or more cells in the subject with a fusion protein that includes a J domain protein and a targeting molecule, wherein the targeting molecule binds the biomolecule. In one aspect, the biomolecule can be a target protein that may be mutated and / or include one or more intrinsically disordered regions. In another aspect, the targeting molecule can be a nanobody, but other targeting molecules are also contemplated. In still another aspect, the disclosed method is useful for treating and / or preventing cancers such as blood cancers and non-small cell lung cancer. Also disclosed are methods for disrupting condensates in cell culture.

The application discloses a stapled peptide compound for selectively degrading MYC protein, a preparation method and application thereof; the compound is sequentially connected in the N-terminal to C-terminal direction by a peptide recognition segment, a flexible linker and a chaperone-mediated autophagy recognition motif KFERQ or a similar motif; the peptide recognition segment is introduced by an (i, i+7) site alkenyl amino acid and a Grubbs catalytic RCM reaction to form a full carbon hydrogen stapling bridge to stabilize the alpha-helix conformation; stapled modification of a typical compound MAX-7 significantly improves the alpha-helix content, the protease stability and the cell penetration, so that the compound can efficiently enter cells and be located in lysosomes, and MYC protein is selectively degraded through the CMA pathway; in-vitro experiments show that MAX-7 can inhibit the proliferation, migration and invasion of various MYC-driven tumor cells, and induce apoptosis; the application provides a brand-new lysosome-directed degradation strategy for the "undruggable" target MYC, and establishes a universal technical platform which can be popularized to other difficult drug proteins.

The present disclosure relates to engineered phospholipid or lysophospholipid (e.g., sphingosine 1-phosphate (S1P)) chaperones derived from apolipoprotein M (ApoM)-Fc fusion proteins with extended half-life in vivo. The disclosed ApoM-Fc fusion proteins provide a safe, highly efficient and effective way of delivering S1P to endothelial cells and all tissues of the body.

The present disclosure relates to methods of producing and screening libraries of disulfide rich proteins, for instance to identify binding peptides specific for a target molecule. In some embodiments, the binding peptides comprise disulfide rich proteins. Also provided herein are methods of producing and screening libraries comprising disulfide-rich proteins and molecular chaperones. The present disclosure also relates to methods of producing or expressing soluble disulfide rich proteins, for instance using a suitable host cell. Also provided herein are compositions comprising soluble disulfide rich proteins.

In one aspect, the invention provides a peptide comprising a chaperone-mediated autophagy (CMA)-targeting signal domain; a protein-binding domain that selectively binds to a target cytosolic protein; and a cell membrane penetrating domain (CMPD). In another aspect, the invention provides methods for reducing the intracellular expression level of an endogenous target protein in vitro and in an animal, wherein the method involves administration of the peptide. Methods are also provided for treating a pathological condition in an animal, the methods comprising administering the peptide to the animal. In one embodiment, the pathological condition is a neurodegenerative disease. In another embodiment of the invention, the target cytosolic protein is death associated protein kinase 1 and the CMPD is protein transduction domain of the HIV-1 Tat protein.

Provided are chaperone retention effect drug conjugates (CREDCs) comprised of an exatecan-like portion, a linker, and a chemical moiety that binds HSP90 protein. Pharmaceutical compositions comprising the disclosed CREDCs and their use in treating cancer and related conditions are also provided.

The invention discloses a recombinant prokaryotic expression vector, an expression strain, soluble human-derived protein and application, the recombinant prokaryotic expression vector comprises a first nucleotide sequence for coding a target human-derived protein and a second nucleotide sequence for coding a cold-adapted molecular chaperone, and a rare codon corresponding to host bacteria is introduced into the first nucleotide sequence to obtain a recombinant prokaryotic expression vector. The translation rate of a key site is actively reduced, a cold-adapted molecular chaperone is introduced into a second nucleic acid sequence, and correct folding and soluble expression of the human-derived protein in a prokaryotic host are promoted in a low-temperature induction manner, so that the high-yield and high-solubility human-derived soluble protein is obtained. The invention provides a feasible way for large-scale preparation of human soluble protein, and has important industrial application value.

The invention discloses a novel molecular chaperone-mediated autophagy (CMA) activating agent, relates to the technical field of biological cells, and is characterized in that the novel molecular chaperone-mediated autophagy activating agent is a novel anti-tumor drug chidamide, and the novel molecular chaperone-mediated autophagy activating agent is a novel anti-tumor drug chidamide. The chidamide can obviously up-regulate the mRNA and protein level of the key protein LAMP2A in the CMA process. According to the present invention, by constructing a human THP1-LAMP2luciferase reporter gene system, the compound capable of activating the LAMP2 promoter is screened from the bulk drug; through cell level primary screening, secondary screening and animal level verification, an effective CMA activator chidamide is finally determined. A good tool is provided for basic research of CMA, and novel drugs are further researched and developed by taking CMA as a target spot.

The invention relates to a method for improving the prokaryotic expression quantity of CRISPR-dCas9 (clustered regularly interspaced short palindromic repeats) protein. Specifically, the invention relates to a method for improving the expression level of dCas9 protein in a prokaryotic cell, and a leucine zipper gene sequence and a dCas9 gene sequence are connected and introduced into the prokaryotic cell. According to the method, the high-purity and high-concentration dCas9 protein can be obtained, the expression quantity of the dCas9 protein is remarkably higher than that of a common method using maltose binding protein (MBP) as a fusion molecular chaperone, the obtained dCas9-lzip fusion protein still has the capability of being normally combined with DNA in a targeted manner under the guidance of sgRNA, and the function of dCas9 is not influenced by connection of a lzip sequence.

The application discloses preparation and application of a heat-sensing repair type recombinant collagen, and relates to the technical field of bioengineering and biomedical materials.The application innovatively carries out gene fusion on a functional domain fragment of a type I humanized collagen and a functional domain fragment of a heat shock protein family through a flexible linker.The recombinant protein is used for solving problems of protein denaturation, membrane structure damage, release of inflammatory factors, impairment of skin barrier function, aggravation of water loss and sensitivity and vulnerability of skin caused by photothermal damage of skin under photothermal stress conditions (such as ultraviolet radiation, high-temperature environment or medical and beauty photoelectric treatment of thermal MAGE, etc.), and the HSP molecular chaperone function is used for guiding damaged proteins to correctly fold and maintain intracellular homeostasis, and the collagen part is used for accelerating tissue repair.Experiments prove that the recombinant protein can effectively improve skin tightness, anti-wrinkle property and barrier repair capacity after thermal damage.The application can be widely applied to the fields of cosmetics, skin repair and protection and medical and beauty postoperative care, etc.

The present invention provides compositions and methods for treating subjects with SCAP-related disorders. [Solution] The present invention provides a double-stranded ribonucleic acid (dsRNAi) agent and composition that targets the SCAP gene, as well as a method for inhibiting the expression of the SCAP gene using such a dsRNAi agent and composition, and a method for treating subjects with SCAP-related disorders such as non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

Provided are a protein for increasing the expression yield of an exogenous protein and the use thereof, wherein an ERV29 protein and a related protein, and a P180 protein and a related protein can be used as molecular chaperones to improve the expression yield of exogenous proteins such as brazzein, the ERV29 protein has an amino acid sequence as shown in SEQ ID No. 4, and the P180 protein has an amino acid sequence as shown in SEQ ID No. 6.

The invention discloses a preparation method and application of high-temperature-resistant and high-pressure-resistant stable yolk microgel, and belongs to the technical field of food processing. The inherent structure of yolk is destroyed through step-by-step hydrolysis of double enzymes, then the protective effect of a molecular chaperone is utilized, and heat-induced self-assembly is combined, so that microgel particles which are compact in structure and uniform in dispersion are constructed, and the particles can effectively resist high-temperature and high-pressure treatment and keep the long-term stability of a system; the method has a wide application prospect in high-protein beverages, liquid nourishment and other liquid foods needing high-temperature sterilization.

Provided herein are chaperone retention effect drug conjugates (CREDCs) comprising an etiotecan-like moiety, a linker, and a chemical moiety that binds to the HSP90 protein. Also provided herein are pharmaceutical compositions comprising the disclosed CREDCs and their use for the treatment of cancer and related conditions.

The application belongs to the technical field of genetic engineering and microbial metabolic engineering, and particularly relates to a modular construction method of a high-titer LanM protein engineering strain. The construction method realizes high-level soluble expression of LanM protein in Escherichia coli through four-dimensional modular synergistic regulation of spatial positioning optimization, chaperone-assisted folding, central carbon metabolic flow redistribution and amino acid synthesis strengthening. The system not only improves the yield of the target protein, but also provides a technical reference for the heterologous expression of other metal-binding proteins with similar structural characteristics. Through the synergy of plasmid copy number control, promoter strength matching and induction timing between the modules, the dynamic balance of intracellular metabolic flow and protein synthesis flow is achieved, meeting the requirements of industrial large-scale production.

The present disclosure provides systems, methods, and kits for directly calibrating fluorescence sensitivity in a flow cytometer using lipid-enveloped droplets comprising first, second, and third known concentrations of an analyte encapsulated in a lipid-enveloped droplet, and wherein the analyte has a fluorescent dye. In embodiments, an analyte encapsulated in a droplet of a lipid envelope includes a fluorescent dye conjugated to a desoxyribonucleic acid (DNA), a ribonucleic acid (RNA), a hydrophilic chaperone that can prevent the analyte from passing through the lipid envelope of the droplet, a protein, an antibody, an antibody fragment, a nanoantibody, or a combination thereof.

The invention discloses preparation and application of thermal induction repair type recombinant collagen, and relates to the technical field of bioengineering and biomedical materials. The functional domain fragment of the I-type humanized collagen and the functional domain fragment of the heat shock protein family are creatively subjected to gene fusion through a flexible connexon. The recombinant protein is used for solving the problems of protein denaturation, membrane structure damage, inflammatory factor release, skin barrier function damage, aggravated moisture loss, sensitivity and fragility and the like caused by photo-thermal damage of skin under photo-thermal stress conditions (such as ultraviolet radiation, high-temperature environment or medical beauty photoelectric therapy such as thermal Margari), and the HSP molecular chaperone function is used for preparing the recombinant protein. The damaged protein is correctly folded, the intracellular homeostasis is maintained, and the tissue repair is accelerated in cooperation with the collagen part. Experiments prove that the recombinant protein can effectively improve skin compactness, wrinkle resistance and barrier repair ability after thermal damage. The invention can be widely applied to the fields of cosmetics, skin repair, post-medical art care and the like.

The disclosure herein provides a fusion protein comprising a prolyl 4-hydroxylase alpha subunit (P4HA) and a soluble protein chaperone. A fusion protein comprising a prolyl 4-hydroxylase alpha subunit (P4HAl) and a prolyl 4-hydroxylase beta subunit (P4HB) is provided. A microorganism comprising a fusion protein comprising a prolyl 4-hydroxylase alpha subunit-1 (P4HAl) and a prolyl 4-hydroxylase beta subunit (P4HB) is provided. The disclosure provides a microorganism comprising a fusion protein comprising a prolyl 4-hydroxylase alpha subunit-1 (P4HAl) and a prolyl 4-hydroxylase beta subunit (P4HB); and another protein to be hydroxylated. A method for providing a skin care benefit comprising applying a fusion protein of the disclosure to the skin is also presented.

The present disclosure relates to methods of producing and screening libraries of disulfide rich proteins, for instance to identify binding peptides specific for a target molecule. In some embodiments, the binding peptides comprise disulfide rich proteins. Also provided herein are methods of producing and screening libraries comprising disulfide-rich proteins and molecular chaperones. The present disclosure also relates to methods of producing or expressing soluble disulfide rich proteins, for instance using a suitable host cell. Also provided herein are compositions comprising soluble disulfide rich proteins.

The application discloses application of a corn ZmRbcL_n1 gene in improvement of photosynthesis efficiency of plants and relates to the technical field of plant genetic engineering. Specifically, the protein coded by the corn ZmRbcL_n1 gene is different from the traditional RbcL protein structure, the protein structure has a molecular chaperone domain for promoting complex assembly, and fluorescence quantification shows that the corn ZmRbcL_n1 gene is highly expressed in leaves, which indicates that the protein may promote photosynthetic carbon fixation of leaves by mechanisms such as promoting RuBisco complex assembly. Further, the application constructs an expression vector for over-expressing the corn ZmRbcL_n1 gene, and the gene is introduced into Arabidopsis thaliana plants by an agrobacterium-mediated method, and it is found that over-expression of the gene can promote biomass accumulation and seed yield of the Arabidopsis thaliana, so that it is indicated that the corn ZmRbcL_n1 gene has great application prospect in improving photosynthesis and increasing carbon fixation efficiency of plants.

The invention belongs to the technical field of recombinant biology, in particular to the field of protein expression. The present invention relates generally to a method of increasing the yield and / or titer of at least one protein of interest in a eukaryotic host cell, preferably yeast, comprising overexpressing in said host cell a) at least one polynucleotide encoding at least one Sbh1 transporter, and b) (i) at least one polynucleotide encoding at least one Kar2 endoplasmic reticulum helper protein; or (ii) at least one polynucleotide encoding at least one Hac1 transcription factor. The invention also relates to a recombinant eukaryotic host cell for preparing at least one protein of interest, engineered to overexpress at least one polynucleotide encoding said at least one Sbh1 transporter, and b) (i) at least one polynucleotide encoding said at least one Kar2 endoplasmic reticulum helper protein; or (ii) at least one polynucleotide encoding the at least one Hac1 transcription factor, and the use of the recombinant eukaryotic host cell for the preparation of at least one protein of interest.

Provided are chaperone retention effect drug conjugates (CREDCs) comprised of an exatecan-like portion, a linker, and a chemical moiety that binds HSP90 protein. Pharmaceutical compositions comprising the disclosed CREDCs and their use in treating cancer and related conditions are also provided.

The present invention relates to a protein composition of Arabidopsis thaliana propilin 2 (AtPFN2) that acts as a molecular chaperone under heat shock stress. More specifically, the invention relates to a protein composition of Arabidopsis thaliana propilin 2 (AtPFN2) that protects a plant from heat shock by acting as a molecular chaperone under heat shock stress and converting into a high molecular weight complex.

The application discloses a sweet protein secretion expression system and a construction method and application thereof, belongs to the technical field of genetic engineering, and specifically discloses an engineered bacterium for expressing thaumatin, an expression vector, a molecular chaperone and an expression method. The thaumatin expressed by the engineered bacterium has a sweet taste after artificial trial, has higher purity and thermal stability compared with natural extraction thaumatin from African canes, the biosynthesis method is more moderate than a chemical extraction method, has higher yield, and is suitable for industrialization.

The application discloses an HSC70 protein K128R mutant and application thereof in regulating CMA activity. The application discloses application of ubiquitination modification of an HSC70 protein at a K128 site in regulating chaperone-mediated autophagy. It is found for the first time that the HSC70 protein is subjected to K11-connected polyubiquitination under metabolic stress, and the modification site is a lysine at the 128th position (K128). The modification is mediated by a ROS-ATM-Atg7-TRIM21 signal axis, and is a necessary condition for the combination of the HSC70 and a lysosome membrane receptor LAMP2A to be enhanced and CMA to be activated. By constructing a K128R mutant, it is proved that the modification loss can block the response of CMA to metabolic stress. The application provides a new diagnostic marker and a drug target for CMA-related diseases (such as tumors and neurodegenerative diseases), and has important clinical application value.