Introduction of Artificial Allosteric Sites on Functional Proteins to Externally Control Function and Binding
Functional proteins, used in this context, are signal amplifiers since interaction with one molecule of the modulator (designed or natural) leads to binding to, or catalyzing the reaction of, more than one molecule of substrates. The read-out can be a part of any well-established reporting system, such as are used e.g. in ELISA systems. The chimera can be a surface-bound or a solution probe.
Amplified molecular detection of a molecule, such as a specific DNA sequence, which in this invention is the modulator;Investigation of protein structure-function relation; Development of "smart drugs"
The present invention involves a method of artificially introducing an allosteric site into a protein whereby an addition of an artificially designed or a natural modulator of that allosteric site leads to a mechanical tension onto the protein. This mechanical tension alters the proteins affinity for its substrate, and, in the case of an enzyme, the catalysis rate.This differential change in substrate affinity or catalysis rate caused by modulator binding can be used in multiple applications. One is the use of such chimera system as an amplified molecular probe, to detect the presence of a natural biological modulator specific to the chimera in an in vitro or in vivo assay. In addition, the chimera-modulator system can be used as a method to study protein conformation and the effect of modulating such conformation on a proteins function.
Patent Number: US8008053B2
Application Number: US2007814275A
Inventor: Zocchi, Giovanni | Choi, Brian
Priority Date: 20 Jan 2005
Priority Number: US8008053B2
Application Date: 18 Jul 2007
Publication Date: 30 Aug 2011
IPC Current: C07K001400 | C07H002100 | C12N000912 | C12Q000148 | C12Q000168
US Class: 435194 | 43500618 | 435015 | 530350 | 5360231 | 435006
Assignee Applicant: The Regents of the University of California
Title: Allosteric control of proteins by manipulating mechanical tension
Usefulness: Allosteric control of proteins by manipulating mechanical tension
Summary: The method is used for altering the conformation of the polypeptide having a known three-dimensional structure. The composition is useful for detecting the presence of a target molecule in a sample, which involves contacting the sample with a composition comprising a polypeptide coupled to a synthetic polymer, where the target molecule is the chemical signal, and detecting the presence of the detectable signal, where presence of the detectable signal is indicative of the presence of the target molecule (claimed). The method is used for designing drugs, treating tumor and constructing amplified molecular probes.
Novelty: Altering conformation of a polypeptide having known 3-dimensional structure, comprises attaching first end and second end of polymer to first and second portion of polypeptide, and altering mechanical tension of polymer
Disease Diagnostic/Treatment
Cancer/Tumor
8008053
State Of Development The following proof-of-concept experiments have been performed: 1. Engineering, expression, and purification of a protein with an artificial allosteric site; 2. Comparison of substrate affinity of the chimera protein versus the native protein; 3. Demonstration of change in binding affinity to substrate upon introduction of a designed modulator; 4. Comparison of different forms of a chimera protein to identify one with the biggest change in substrate affinity upon modulator introduction. Next-steps: Generation of a other chimeras, using different allosteric proteins and corresponding substrates, to generate a chimera that:a. Demonstrates a substantial change in substrate binding or catalysis rate upon introduction of modulator;b. Demonstrates activity to a reliable, easy-to-use, and well-studied reporter system. Related Materials Allosteric control through mechanical tension. Phys Rev Lett. (2005) Tech ID/UC Case 20197/2004-487-0 Related Cases 2004-487-0
Mechanical control of Renilla Luciferase. J Am Chem Soc. (2013)
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