The General Hospital Corporation

Other

Harvard College

Genetics Institute, Inc.

The General Hosptial Corporation

The Molecular Sciences Institute, Inc.

Roger Brent

Antonis S Zervos

John M McCoy

Erica A Golemis

Timm H Jessen

Andrew R Mendelsohn

Walter L Lok

Russell L Finley, Jr

C Wilson Xu

Jeno Gyuris

Mark S Ptashne

Robert H Carlson

Chanxing Wilson Xu

Catherine Anderson

Karen F Lech

Ian E Burbulis

Disclosed herein are methods for detecting complex protein interactions and protein functional relationships, and reagents for carrying out those methods.

Detection systems for registering protein interactions and functional relationships

A system and method for characterizing protein molecules. A protein molecule of interest is isolated from other types of protein molecules. The protein molecule of interest is modified to a one-dimensional structure from the natural three-dimensional structure of the protein molecule. Each of a first type of amino acid residue of the protein molecule is labeled with a first tag. Each of a second type of amino acid residue of the protein molecule is labeled with a second tag. The first and second tags impart to the protein molecule a detectable set of characteristic ancillary properties that facilitates distinction of the protein molecule of interest from other types of protein molecules. When these ancillary properties are detected, a fingerprint of the protein molecule is revealed. A listing of known protein molecules and of the fingerprints corresponding to each of the known protein molecules serves as a library to facilitate identification of unknown proteins. A fingerprint of a protein molecule of interest determined in the same manner as the fingerprints of known proteins listed in the library is compared with fingerprints of the known protein molecules to identify the protein molecule of interest.

Protein fingerprint system and related methods

Disclosed herein is a method of determining whether a first protein is capable of physically interacting with a second protein, involving: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a gene activating moiety and being conformationally-constrained; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Also disclosed are methods for assaying protein interactions, and identifying antagonists and agonists of protein interactions.

Interaction trap systems for detecting protein interactions

Disclosed herein is a method of determining whether a first protein is capable of physically interacting with a second protein, involving: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a gene activating moiety and being conformationally-constrained; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Also disclosed are methods for assaying protein interactions, and identifying antagonists and agonists of protein interactions. Finally, populations of eukaryotic cells are disclosed, each cell having a recombinant DNA molecule encoding a conformationally-constrained intracellular peptide.

Disclosed are sets of DNA molecules, and cell containing such molecules, each molecule encoding a candidate interacting protein fused to either a DNA binding domain or a gene activating domain to which it is not naturally bonded.

Interaction trap systems for analysis of protein networks

Disclosed are substatially pure Reparations of Max-Interacting (Mxi) polypeptides, DNA encoding such polypeptides, antibodies recognizing such polypeptides, and diagnostic and therapeutic methods utilizing such polypeptides.

Methods of detecting a malignant cell in a biological sample comprising

Disclosed herein is a method of determining whether a first protein is capable of physically interacting with a second protein, involving: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a gene activating moiety and being conformationally-constrained; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Also disclosed are methods for assaying protein interactions, and identifying antagonists and agonists of protein interactions. Proteins isolated by these methods are also discussed. Finally, populations of eukaryotic cells are disclosed, each cell having a recombinant DNA molecule encoding a conformationally-constrained intracellular peptide.

Disclosed are substatially pure preparations of Max-Interacting (Mxi) polypeptides, DNA encoding such polypeptides, antibodies recognizing such polypeptides, and diagnostic and therapeutic methods utilizing such polypeptides.

Max-interacting proteins and related molecules and methods

Disclosed is a method for determining whether a first protein is capable of physically interacting with a second protein. The method involves: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a weak gene activating moiety; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Such a determination facilitates the isolation of the gene encoding the interacting protein. Also disclosed herein is recombinant Cdi1 polypeptide, nucleic acid encoding the Cdi1 polypeptide, and uses thereof.

CDI1 polypeptides

Disclosed are improved methods for detecting protein-protein interactions. These methods involve either a determination of whether three or more proteins are capable of interacting in a trimeric or higher order complex or whether two or more mammalian proteins interact, the latter method utilizing yeast mating to bring candidate proteins into contact with one another.

Assays for inhibitors of myc oncoprotein

Interaction trap system for isolating novel proteins

Disclosed are substantially pure preparations of Max-Interacting (Mxi) polypeptides, DNA encoding such polypeptides, antibodies recognizing such polypeptides, and diagnostic and therapeutic methods utilizing such polypeptides.

Growing community of inventors

Cambridge, MA, United States of America

Roger Brent