49 results about "Phosphorylation site" patented technology

The invention discloses a tree shrew Alzheimer's disease modeling method. The method comprises the following specific steps: mixing an ethanol solution of Microcystin-LR with the concentration of 0.5 [mu] g / [mu] l with an amaninic acid solution, and injecting 2 [mu] l to Meynert basal nuclei on the two sides of a tree shrew respectively; and after drug administration is finished, normally feeding for 2 weeks to finish Alzheimer's disease modeling. The method comprises the following steps: under a sterile condition, sucking 200 microliters of sterile normal saline, adding the sterile normal saline into a small bottle containing 1mg of amanitine, repeatedly blowing and beating to completely dissolve the amanitine, and preparing a solution with the concentration of 5mu g / microliter; the Microcystin-LR solution is a product purchased in the market. According to the invention, a mixed solution of MC-LR (0.1 [mu] g) and IBO solution (3 [mu] g) is respectively injected to Meynert basal nuclei on both sides of the tree shrew, and a stable tree shrew AD model can be established after 2 weeks. After the medicine is administrated for 3 days, tree shrews have AD behavior expressions, such as drooping spirit and reduced activity; after two weeks, the tree shrew brain shows AD typical pathological expressions such as A beta deposition, increase of expression levels of multiple phosphorylation sites of Tau protein and the like.

The invention discloses annexin A2 (ANXA2) and application of an inhibitor thereof in diagnosis, treatment and / or prevention of pulmonary arterial hypertension. In particular discloses application of an ANXA2 inhibitor (including siRNA for silencing an ANXA2 gene, an ANXA2 antibody and a phosphorylation inhibitor) in preparation of a product for preventing and / or treating pulmonary arterial hypertension. Experiments prove that the ANXA2 inhibitor can significantly inhibit proliferation and migration of pulmonary artery smooth muscle cells, and significantly improve pulmonary arterial hypertension, pulmonary artery vascular remodeling and right ventricular hypertrophy. The ANXA2 or the Thr208 phosphorylation site of the ANXA2 protein can be used for diagnosis or auxiliary diagnosis of pulmonary arterial hypertension, or used for screening drugs for pulmonary arterial hypertension and developing new diagnosis and treatment methods and drugs. The therapeutic target and the ANXA2 inhibitor developed by the invention have wide clinical application value in the fields of diagnosis, prevention and treatment of pulmonary arterial hypertension.

The invention relates to the technical field of biological information, and discloses a multi-mode phosphorylation site prediction method and device, equipment and a medium, and the method comprises the steps: extracting a subsequence of a candidate phosphorylation site from a to-be-predicted protein sequence, and generating a sequence feature of the subsequence through a deep learning model; constructing a graph structure based on the three-dimensional structure information of the candidate phosphorylation sites, and extracting structural features of the candidate sites through a graph neural network according to the graph structure; performing multi-modal feature fusion on the sequence features and the structural features to generate fusion features; and predicting the phosphorylation probability of the candidate site based on the fusion feature to obtain a phosphorylation probability prediction result. According to the method, the protein sequence language model and the multi-modal features of the three-dimensional structure diagram convolutional network are fused, so that the accuracy and comprehensiveness of phosphorylation site prediction can be effectively improved, and meanwhile, the calculation efficiency and the bioinformatics interpretation are considered.

The invention belongs to the technical field of biological materials and biological information, and provides a space phosphorylation modification omics detection method. According to the detection method disclosed by the invention, proteomics analysis can be carried out on trace sample phosphorylation, 4259 phosphorylation sites can be identified by 5 micrograms of peptide fragments, and the credibility of 3506 sites is greater than 0.75.

The invention discloses application of an NR1D1 protein phosphorylation site in preparation of cerebral hemorrhage treatment drugs, and belongs to the technical field of molecular biology. According to the invention, the specific phosphorylation site and phosphorylation modification level of the NR1D1 protein are determined for the first time, and the NR1D1 protein can be used as a key marker for evaluating the secondary nerve injury degree of the hemorrhagic cerebral apoplexy and developing treatment and / or alleviation of the hemorrhagic cerebral apoplexy; a specific antigen peptide and an antibody for accurately detecting the NR1D1 protein phosphorylation site are further constructed, and an efficient tool is provided for molecular evaluation of hemorrhagic stroke secondary nerve injury; meanwhile, polypeptide molecules capable of inhibiting NR1D1 protein phosphorylation are researched and developed, through the dual effects of specifically blocking the phosphorylation process and reducing protein degradation, necrosis of brain tissues around hematoma after hemorrhagic cerebral apoplexy is relieved, secondary dyskinesia is improved, and the effect of inhibiting NR1D1 protein phosphorylation is achieved. And a brand new scheme is provided for molecular evaluation and targeted intervention of hemorrhagic stroke secondary nerve injury.

The invention discloses a protein sequence phosphorylation site prediction method and system, computer equipment and a storage medium, and relates to the technical field of protein sequence phosphorylation site prediction.The protein sequence phosphorylation site prediction method includes the steps that data collection is conducted on an amino acid sequence of a protein and a corresponding phosphorylation site, preprocessing is conducted in an automatic labeling mode, and a prediction result is obtained; dividing the protein sequence into a plurality of polypeptide chains through sliding window calculation; the method comprises the following steps: coding an amino acid sequence based on a BERT protein language model, extracting protein information features, calculating an emission score and a transfer score by taking a conditional random field (CRF) as a decoder, capturing regional distribution and a tandem effect of phosphorylation sites, and carrying out loss calculation by taking a negative likelihood logarithm (NNL) as a loss function, protBERT is adopted as a main framework for extracting protein sequence information to carry out establishment and training of a phosphorylation prediction model; and carrying out phosphorylation prediction through the phosphorylation prediction model. According to the method, the labor cost and the time cost are greatly reduced.

The application discloses an antigen peptide targeting DRP1 protein Ser572 phosphorylation, an antibody and application thereof, and particularly relates to the technical field of biological medicines, and the antigen peptide has immunogenicity of targeting a DRP1 protein Ser572 phosphorylation site, wherein the amino acid sequence of the antigen peptide is RETKNVA{piSer}GGGGVGC, and {piSer} indicates that serine at the site is modified by phosphorylation. The antibody prepared by the application can stably recognize DRP1 Ser572 phosphorylation modification at a cell and animal level, can clearly reveal a molecular mechanism of the site phosphorylation in regulating DRP1 liquid-liquid phase separation and mediating myocardial mitochondrial damage, can support basic mechanism research of diseases such as sepsis, can be used for developing a specific ELISA kit and a pharmaceutical composition, fills a technical gap of DRP1 Ser572 phosphorylation related research and clinical transformation, and provides a brand-new technical support for diagnosis and treatment of cardiovascular diseases, sepsis and other major diseases.

The invention relates to an application of a zinc finger protein Zfp260 phosphorylation site for regulating osteogenesis, and particularly discloses an application of a biomarker in preparation of a medicine for regulating bone regeneration and repair, and the biomarker comprises one or a combination of more of Y173 phosphorylation, S182 phosphorylation and S197 phosphorylation of the zinc finger protein Zfp260. The invention finds that any phosphorylation defect mutation of the three sites Y173, S182 and S197 can obviously change the binding force and nuclear translocation signal with kinase PKC alpha, and each mutant cannot save the cell mineralization defect caused by Zfp260 knockout, and prompts that the newly found Zfp260 phosphorylation sites Y173, S182 and S197 play important roles in Zfp260 related diseases and the regulation process of promoting bone regeneration and repair.

The invention discloses an IBDV (Infectious Bursal Disease Virus) mutant based on VP1 protein T425 and R426 synergistic modification and application of the IBDV mutant. The invention identifies that threonine at the 425th site on the VP1 protein is a new phosphorylation site for the first time, and reveals that threonine at the 425th site and adjacent arginine methylation modification at the 426th site have a synergistic positive feedback regulation relationship. Based on the mechanism, a T425 and R426 double-site modification deleted virus mutant is successfully constructed, and the mutant shows a phenotype with severely weakened replication ability. In addition, the invention also provides an antibody capable of specifically recognizing pT425, an application of a GSK-3 kinase inhibitor in inhibition of virus replication, and a related screening and detection method. The invention provides a new target and strategy for developing novel IBDV attenuated vaccines, antiviral drugs and diagnostic reagents.

This invention provides a fluorescent probe for label-free, multifunctional, low-cost, and high-throughput real-time monitoring of tyrosine phosphorylation. Our designed fluorescent molecule specifically binds to copper ions in aqueous solution, leading to fluorescence quenching. Adding a phosphorylated substrate peptide with tyrosine residues to the quenched system elicits a dramatic change in fluorescence, with the solution color changing from purple to red. However, no significant fluorescence change is observed upon adding non-phosphorylated substrate peptides or other phosphate-related ions. This on-off fluorescence response enables real-time monitoring of tyrosine phosphorylation, with a detection limit of 100 nM. This method can also be applied to the rapid screening of PKA kinase inhibitors and to the detection of serine, threonine, and phosphorylated peptides at mono- and diphosphorylated sites. The greatest advantage of this fluorescence spectroscopy method is that it does not require antibodies or radioactive isotopes, has simple operating procedures, and is highly suitable for the real-time detection of protein tyrosine phosphatase activity.

According to the application of the reagent for detecting the phosphorylation level of the S202 and T205 sites of the MAPT protein in preparation of the renal fibrosis diagnosis product, the function research of the MAPT protein is expanded from the traditional field of glomerular podocytes to renal tubular epithelial cells and the fibrosis pathological process of the renal tubular epithelial cells for the first time, and a brand new direction of the MAPT protein in renal disease research is opened up. Different from the conventional general research means which only depends on overall gene knockout or overexpression, the method provided by the invention realizes accurate analysis of the function of the MAPT key phosphorylation site by using the site-specific mutant. Through a systematic function determination experiment, the invention discloses an anti-intuition biological rule with great theoretical significance: although phosphorylation of the sites S202 and T205 obviously drives the fibrosis process of the renal tubular epithelial cells, the fibrosis phenotype cannot be improved by simulating the dephosphorylation state (S202A / T205A mutant) of the sites.

This application relates to the field of biomedical technologies, and specifically discloses a transcription factor HOMEZ phosphorylation site-specific antibody, its preparation method and applications. The specific antibody provided in this application can specifically recognize the phosphorylation of threonine at position 451 of HOMEZ in tumor specimens, thereby judging the prognosis of tumors and providing potential therapeutic targets for the diagnosis and treatment of clinical tumors. Moreover, the specific antibody of this application can be used in Western blot experiments to detect the expression differences between normal cells and tumor cells, which helps to study the role of phosphorylation modification of HOMEZ protein in the occurrence and development of tumor diseases. It can also be used in experiments such as immunofluorescence and immunohistochemical staining to detect the phosphorylation level of HOMEZ protein in clinical specimens, and has broad clinical application prospects in aspects such as disease diagnosis, treatment and prognosis determination.

The invention discloses TAL1 and TAL1S122A mutant recombinant proteins as well as a preparation method and application of the TAL1 and TAL1S122A mutant recombinant proteins. The amino acid sequence of the TAL1 and TAL1S122A mutant recombinant protein provided by the invention is as shown in SEQ ID NO: 1 or SEQ ID NO: 2. The recombinant protein can be used in in-vivo and in-vitro experiments of phosphorylation of TAL1 by MAP3K3 and other kinases. The invention also provides a preparation method of the recombinant protein. The preparation method comprises the steps of expression plasmid construction, expression test, amplification expression and affinity purification, protein dialysis, protein concentration and the like. The concentrated recombinant protein has the characteristics of high purity and good activity. In an in-vitro kinase experiment with a commercial MAP3K3 recombinant protein, the TAL1 recombinant protein can be smoothly phosphorylated by MAP3K3, and the phosphorylation degree of S122A mutation by MAP3K3 is reduced. The application has important significance for researching the TAL1 and the phosphorylation site of the TAL1.

This invention relates to the field of pharmaceutical technology for treating diseases related to heavy metal exposure, specifically to the application of a small molecule compound targeting the phosphorylated serine 43 site of ARMC10 in the preparation of drugs for treating neurological damage caused by tin exposure. This technical solution is the first to discover that trimethyltin chloride exposure can specifically induce abnormal phosphorylation of ARMC10, thereby causing neuronal death and cognitive impairment. Based on this mechanism, a small molecule compound targeting a specific phosphorylation site of ARMC10 is proposed, which effectively reverses neurological damage and protects neuronal structure and function by blocking its mediated abnormal mitochondrial division and dysfunction. This solution solves the problem of existing technologies lacking effective treatments for tin exposure-related neurological damage due to the lack of clear drug targets, and has ideal application prospects. This small molecule compound has a well-defined structure and a clear mechanism of action, is easy to further develop, and possesses good drug development and commercialization prospects.

The invention discloses a targeted ATAD3A phosphorylated polypeptide and application thereof in preparation of an anti-aging product. The targeted ATAD3A phosphorylated polypeptide is composed of a functional peptide with an amino acid sequence as shown in SEQ ID No.1 and a cell-penetrating peptide connected with the N end of the functional peptide. Research finds that overexpression of phosphorylated ATAD3A promotes cell aging, which prompts that targeted inhibition of ATAD3A phosphorylation may have an anti-aging effect. Therefore, the invention designs the TAT-polypeptide targeting ATAD3A phosphorylation site for anti-aging treatment, and the TAT-polypeptide shows a good treatment effect in both cellular level and mouse anti-aging treatment. Therefore, the targeted ATAD3A phosphorylated polypeptide opens a new thought and provides a new target for exploring anti-aging treatment, and the polypeptide can be further used for preparing anti-aging drugs and relieving the pressure brought by population aging to social medical care.

The invention designs a biosynthetic hydroxyapatite-based protein kinase B (Akt1) electrochemical sensor which is applied to Akt1 activity analysis in a skin cell aging process. Hydroxyapatite is biologically synthesized through bacillus subtilis and is uniformly deposited on the surface of a glassy carbon electrode by utilizing a potentiometric deposition method. The surface of the material contains rich phosphate groups and hydroxyl groups, can be combined with lanthanum ions, and provides a good interface for subsequent sensor detection. In order to detect the activity of Akt1, a substrate polypeptide containing three phosphorylation sites is designed. The Akt1 is subjected to phosphorylation of a substrate polypeptide, negative charges are increased, combination of the Akt1 and lanthanum ions on the surface of the electrode is enhanced, the impedance on the surface of the electrode is increased, and the activity of the Akt1 is reflected. And the unphosphorylated substrate polypeptide cannot be effectively attached and shows a relatively small impedance value. The accuracy of the sensor is verified by adopting an Akt1 inhibitor MK-2206. Human skin fibroblasts (GNHu49) are taken as a model, and research finds that the activity of Akt1 in senescent cells is remarkably reduced, so that an experimental basis is provided for researching the activity change of Akt1 and the change of cell functions in the skin senescence process. The invention provides a new technical means for skin aging research, anti-aging drug evaluation and cosmetic technology.

The invention belongs to the field of molecular biology, and particularly relates to a construction method and application of phosphorylation mutation sites of phosphoglucomutase. The mutation site is obtained by mutating threonine at the 144th site of escherichia coli wild-type phosphoglucomutase into aspartic acid, the sensitivity of an escherichia coli mutant strain with the mutation site to nalidixic acid, mitomycin C and tetracycline is remarkably improved, and the mutation site can be used for improving the sensitivity of bacteria to drugs.

The application provides an antibody identification method, a polypeptide composition, a kit and a preparation method and application thereof, and relates to the technical field of biotechnology. The antibody identification method mainly comprises identifying whether an antibody specifically recognizes a 231st phosphorylation site of Tau protein, reacting a to-be-detected antibody with four identification polypeptides respectively, and determining whether the to-be-identified antibody can specifically recognize the 231st phosphorylation site of Tau protein according to a detection result. The four identification polypeptides are based on a basic polypeptide, and the specificity of the to-be-identified antibody to the target phosphorylation site is determined through the phosphorylation difference between the 231st phosphorylation site and potential phosphorylation sites of Tau protein, so that the problem of a complex antibody screening and identification method for recognizing the 231st phosphorylation site of Tau protein in the prior art is solved.

The invention relates to the field of protein detection, in particular to a phosphorylated peptide fragment preparation and space phosphorylated proteomics detection method. By means of the preparation method, enough phosphorylated peptide fragments can be efficiently extracted from a trace sample, the method can be compatible with LCM for use, operation is easy and convenient, the recovery rate is high, and the method is suitable for achieving high-depth space phosphorylated proteomics analysis. Meanwhile, the analysis method shows high sensitivity on the trace level, and compared with a traditional method, the identification number of phosphorylation sites is remarkably increased. Compared with the homogenization treatment of the whole section, the analysis method provided by the invention can more accurately reflect the specificity of the cells in the target area and better reflect the intra-tumor heterogeneity.

The invention discloses a method for improving activity of aspergillus oryzae amylase through CreA C terminal dephosphorylation mutation, and belongs to the technical field of enzyme engineering. According to the method, a CreA dephosphorylation mutant strain is constructed by performing site-specific mutagenesis on a C-terminal phosphorylation site coding gene of an aspergillus oryzae carbon metabolism inhibition core transcription factor CreA, so that a carbon metabolite repression effect is relieved, and the yield of amylase is increased. Specifically, four phosphorylation sites (S414, S415, T416 and S419) at the C tail end of the CreA protein are mutated into valine (V). When the mutant strain is used for fermentation, the expression of amylase can be remarkably improved. Experiments prove that the amylase activity of the mutant strain 4M can be 3 times that of a reference strain after the mutant strain is cultured for 6 days in a culture medium taking starch as a carbon source. The invention provides a new strain modification strategy for efficient production of amylase.

The invention relates to an application of STAT3 or a phosphorylation inhibitor thereof in prevention or treatment of silent corticotropin adenoma, and particularly provides an application of STAT3 or a protein phosphorylation inhibitor thereof in prevention and / or treatment of silent corticotropin adenoma. According to the invention, it is found for the first time that a signal transduction and transcriptional activation factor 3 (Signal Transduction and Transcriptional Activator of Of Transcriptional Activator 3, STAT3) and a phosphorylation site of the STAT3 have specific enrichment in silent corticotropin adenoma, and the application of the STAT3 to the silent corticotropin adenoma has the advantages that the specificity of the silent corticotropin adenoma and the phosphorylation site of the silent corticotropin adenoma is improved;

The invention relates to the technical field of cell models, in particular to a hepatocyte injury model as well as a construction method and application thereof. The invention provides a model for over-expressing a phosphoethanolamine phosphatase gene ETNPPL to damage liver cell functions, after the ETNPPL is over-expressed in liver cells, reduced glutathione is reduced, and active oxygen, glutamic oxalacetic transaminase and glutamic-pyruvic transaminase are increased; mitochondria vacuolation and structure damage are caused, the number of mitochondria is reduced, and the cell proliferation ability is reduced; the protein expression quantity of Akt phosphorylation sites p-Akt-Thr308 and p-Akt-Ser473 of a signal path PI3K / AKT is reduced, and the PI3K / AKT can be used as a liver cell metabolic function impaired model, is used for analyzing a liver cell injury molecular mechanism and is used as a material for evaluating targeted therapy measures.

The invention belongs to the field of biological medicine, and relates to a separated ubiquitin specific peptidase 30 mutant, nucleic acid, an antibody and application. The ubiquitin specific peptidase 30 mutant is a USP30S104A mutant, and the ubiquitin specific peptidase 30 mutant has an amino acid sequence as shown in SEQ ID NO: 1. The invention discovers a brand new phosphorylation site (104 site) on USP30 protein, and provides a breast cancer gene therapy and chemotherapy drug resistance prediction method based on the mutant and phosphorylation detection by researching a USP30S104A mutant nuclear localization regulation mechanism.

Methods for improving cardiac function are provided. [Solution] Provided is a modified Yes-associated protein (YAP) in which the serine residues at the LATS1 / 2 phosphorylation site and in the region of YAP that binds to TEAD are substituted with alanine. Also provided are nucleic acids encoding the modified YAP, vectors containing the nucleic acid, and compositions containing the modified YAP, as well as nucleic acids encoding the modified YAP or vectors containing the nucleic acid. Also provided are methods for regenerating cardiomyocytes and treating cardiac conditions.

Use of a reagent and / or device for detecting a human ABCG1 phosphorylation site in preparation of a reagent for risk assessment, diagnosis, or prognostic assessment of a tumor, a kit for risk assessment, diagnosis, or prognostic assessment of a tumor, use of a reagent for inhibiting a human ABCG1 phosphorylation site in preparation of a drug for preventing and / or treating a tumor, and a kit for preventing and / or treating a tumor. The human ABCG1 phosphorylation site is one or more selected from the group consisting of Y97, T99, S120, S125, T272, S277, S441, and Y655. A human ABCG1 protein phosphorylation-specific antigenic peptide, an antibody, use of the human ABCG1 protein phosphorylation-specific antigenic peptide or the antibody, and a kit including the human ABCG1 protein phosphorylation-specific antigenic peptide or the antibody.

The invention belongs to the field of immunology, and particularly relates to a monoclonal antibody for recognizing phosphorylation of a Tau protein 181 site and application of the monoclonal antibody. When the monoclonal antibody is prepared, antigen polypeptide designed aiming at the 181st phosphorylation site of Tau protein is adopted, the specific sequence of the antigen polypeptide is PK (pT) PPC, and (pT) represents phosphorylated amino acid. The amino acid sequence of the heavy chain variable region of the monoclonal antibody comprises a CDR1 shown as SEQ ID No. 1, a CDR2 shown as SEQ ID No. 2 and a CDR3 shown as SEQ ID No. 3; the amino acid sequence of the light chain variable region comprises a CDR1 shown as SEQ ID No. 4, a CDR2 shown as SEQ ID No. 5 and a CDR3 shown as SEQ ID No. 6. The monoclonal antibody can be used for carrying out high-specificity, high-sensitivity and high-accuracy detection on the 181 site of the phosphorylated Tau protein.

The invention belongs to the technical field of proteomics, and particularly relates to a phosphorylated peptide fragment enrichment identification method for trace tissues. The method comprises the following steps: (1) adding a lysis solution into a trace tissue sample, and heating; performing ultrasonic treatment on the sample in an ice-water bath; adding enzyme for enzymolysis; adding a solution containing a phosphopeptide enrichment material into the sample for incubation; activating and balancing a peptide fragment desalting column by using two enrichment buffer solutions; (2) adding a sample and a mixed solution containing the phosphopeptide enrichment material into the pretreated desalting column; cleaning with a cleaning buffer solution A and a cleaning buffer solution in sequence; eluting with two elution buffer solutions in sequence; and collecting the eluent, and carrying out vacuum concentration and freeze-drying to obtain the enriched phosphorylated peptide fragment of the trace tissue sample. The method can reduce treatment steps and reduce liquid transfer, so that sample loss can be reduced; the pretreatment time is greatly saved; the identification of the phosphorylation site of the protein with the initial quantity as low as 10 ng is realized.

The present disclosure provides proteins, methods, and systems for targeting enzymes to specific subcellular locations to produce post-translational modifications on proteins. Specifically, kinases for phosphorylating proteins containing one or more sites susceptible to phosphorylation are disclosed, along with methods and systems for phosphorylating proteins of interest, such as proteins with nutritional and therapeutic uses. Recombinant host cells engineered to express a non-naturally occurring polypeptide comprising a heterologous serine / threonine kinase, wherein the heterologous serine / threonine kinase is anchored in an intracellular membrane, are disclosed herein.

The application belongs to the technical field of biological medicine, and relates to a BCLAF1 protein Ser564 site phosphorylation antigen peptide, a specific antibody and a preparation method and application thereof. The application identifies BCLAF1 protein Ser564 as a key phosphorylation site through SIK2 kinase screening, and designs and synthesizes a phosphorylation antigen peptide based on the site. After immunizing animals, a polyclonal antibody specifically recognizing BCLAF1 Ser564 site phosphorylation modification is successfully prepared. The antibody has high sensitivity and strong specificity, is suitable for various detection platforms such as immunohistochemistry and immunoblotting, and can be used for evaluating tumor immunotherapy efficacy. The application provides a key tool for analyzing BCLAF1 phosphorylation regulation mechanism, and lays a theoretical foundation for developing an immune combined therapy strategy targeting a SIK2-BCLAF1-CCL5 axis.

The present disclosure provides for proteins, methods, systems for directing enzymes for generating post-translational modifications on proteins to a particular subcellular location. Specifically, kinases for phosphorylation of proteins containing one or more sites susceptible to phosphorylation are disclosed, also with methods and systems for phosphorylating proteins of interest, such as proteins having nutritional and therapeutic uses.