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    HIGH-THROUGHPUT ENZYME ASSAY FOR SCREENING ACTIVITY OF ACYLTRANSFERASES
    31.
    发明申请
    HIGH-THROUGHPUT ENZYME ASSAY FOR SCREENING ACTIVITY OF ACYLTRANSFERASES 有权

    公开(公告)号：US20220403443A1

    公开(公告)日：2022-12-22

    申请号：US17840140

    申请日：2022-06-14

    申请人： CORNELL UNIVERSITY

    发明人： Hening Lin ,  Maurine E. Linder ,  Jun Young Hong

    IPC分类号： C12Q1/48

    摘要： A method for determining activity of an acyl transferase enzyme, the method comprising: (i) preparing a reaction mixture comprising: (a) an acyl transferase enzyme, (b) a peptide substrate bound to a fluorophore, wherein the substrate is a cysteine-containing oligopeptide of 5-25 amino acids in length, (c) an acyl-CoA, and (d) a detergent comprising micelles, wherein the acyl transferase enzyme mediates acylation on a cysteine of said peptide substrate to result in association of the peptide with micelles of the detergent with resultant increase in fluorescence polarization; and (ii) measuring fluorescent signal of the reaction mixture; wherein an increase in fluorescence polarization of the reaction mixture compared to fluorescence polarization of a control reaction indicates acyl transferase activity of the acyl transferase enzyme. The above assay method may also be used for screening compounds for their ability to act as inhibitors of an acyl transferase enzyme.

    PRENYLATION ASSAY
    33.
    发明申请
    PRENYLATION ASSAY 有权

    公开(公告)号：US20220380831A1

    公开(公告)日：2022-12-01

    申请号：US17559713

    申请日：2021-12-22

    申请人： Oxford University Innovation Limited

    发明人： Robert E. Maclaren ,  Maria Ines Moreira Patricio

    IPC分类号： C12Q1/48

    摘要： A method for determining the activity of Rab escort protein 1 (REP1) comprising the steps: (a) providing a sample comprising REP1; (b) contacting the sample of step (a) with Rab6a, Rab geranylgeranyltransferase (Rab GGTase) and a lipid donor substrate; and (c) detecting the lipidated Rab6a product.

    METHOD FOR CONTROLLING SPEED OF POLYPEPTIDE PASSING THROUGH NANOPORE AND USE THEREOF
    34.
    发明申请
    METHOD FOR CONTROLLING SPEED OF POLYPEPTIDE PASSING THROUGH NANOPORE AND USE THEREOF 有权

    公开(公告)号：US20220365094A1

    公开(公告)日：2022-11-17

    申请号：US17620338

    申请日：2020-06-24

    申请人： TSINGHUA UNIVERSITY

    发明人： Jingwei BAI ,  Zhijie CHEN

    IPC分类号： G01N33/68 ,  G01N27/00 ,  G01N33/558 ,  C12Q1/48 ,  C12Q1/34

    摘要： The present invention provides a method for controlling a speed of a polypeptide passing through a nanopore and use thereof in determining an amino acid sequence of a polypeptide. Specifically, the method comprises: conjugating a polynucleotide to the polypeptide to give a polynucleotide-polypeptide conjugate, and applying a voltage across the nanopore in the presence of a polynucleotide binding enzyme to move the conjugate through the nanopore. The polynucleotide binding enzyme controls the movement of the polynucleotide and thereby controls the movement of the conjugated polypeptide in the nanopore, thus controlling the speed of the polypeptide passing through the nanopore. While controlling the speed of the polypeptide, the present invention reads a nanopore current signal during the process of the polypeptide passing through the nanopore to give an electrical signal of the polypeptide. The electrical signal can be further used to acquire an amino acid sequence of the polypeptide, to identify the polypeptide or a part thereof, or to establish a library of polypeptide electrical signals.

    Methods and composition for modifying enzymes
    35.
    发明授权
    Methods and composition for modifying enzymes 有权

    公开(公告)号：US11459400B2

    公开(公告)日：2022-10-04

    申请号：US15745496

    申请日：2016-07-15

    申请人： The University of Chicago

    发明人： Shohei Koide ,  Shun-Ichi Tanaka ,  Akiko Koide

    IPC分类号： G01N33/53 ,  G01N33/569 ,  G01N33/573 ,  C07K16/40 ,  C12Q1/26 ,  C12Q1/34 ,  C12Q1/48 ,  C12Q1/527 ,  C12Q1/533 ,  C12Q1/00 ,  C12N9/38 ,  C07K14/78 ,  C12N9/20 ,  C12Q1/25 ,  G01N33/58 ,  G01N33/60

    摘要： Aspects of the disclosure relate to compositions of enzyme-binding polypeptides (EBPs) that modify the substrate specificity of an enzyme and a method for identifying an EBP that modifies substrate specificity of an enzyme binding at least one substrate, the method comprising: contacting the enzyme with a polypeptide library comprising a plurality of EBPs that bind different epitopes of the enzyme; identifying EBPs that bind to the enzyme to form an EBP-enzyme complex; assaying for the activity level and substrate specificity of the EBP-enzyme complex; and identifying EBPs that modify the substrate specificity of the enzyme by identifying EBPs that, when in an EBP-enzyme complex, have a different substrate specificity than un-complexed EBP; wherein the catalytic rate constant of the EBP-enzyme complex is ≥50% of the un-complexed enzyme for at least one substrate and/or wherein the EBP-enzyme complex retains binding to a substrate.