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Tandem analyses of noncovalently driven effectors for modulatory mapping of activities of protein sites

  • US 7,927,579 B2
  • Filed: 01/16/2004
  • Issued: 04/19/2011
  • Est. Priority Date: 01/17/2003
  • Status: Active Grant
First Claim
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1. A method for modification of a target group in or near a secondary ligand binding site of a protein comprising:

  • contacting the protein with at least one site-specific activated polymer complex comprising;

    a functional group reactive with the target group;

    a polymer linked to the functional group; and

    a ligand moiety that specifically binds to the secondary ligand binding site, thereby effecting site-specific covalent bonding of the target group on the protein via the functional group,and eliminating the ligand moiety from the protein-polymer conjugate after the specific covalent bonding of the target group on the protein,wherein the functional group is selected from the group consisting of;

    a dithioester;

    a thioloester;

    thionoester;

    a selenoester;

    a selenoloester;

    a selenonooester;

    a phosphonoester;

    a phosphoric ester;

    a phosphinic ester;

    a sulfonate ester;

    a diazoester;

    an acylphosphate;

    an o-salicylate;

    a p-salicylate;

    a m-salicylate;

    a disulfide; and

    an α

    -keto- or β

    -keto acid or ester;

    wherein the polymer is a polyethylene glycol polymer;

    wherein the secondary site-specific ligand moiety is a peptidyl ligand moiety or carbohydrate ligand moiety; and

    wherein the secondary ligand binding site is a site differing from a primary binding site; and

    wherein the secondary ligand binding site has no effect on the primary function of the protein.

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