Allosteric control of proteins by manipulating mechanical tension
AssigneeThe Regents of the University of California
InventorsGiovanni Zocchi, Brian Choi
Life of this patent
Estimating the term…
Abstract
A method of altering the conformation of a polypeptide having a known three-dimensional structure is described. The method comprises attaching a first end of a polymer to a first portion of the polypeptide, attaching a second end of the polymer to a second portion of the polypeptide, and altering the mechanical tension of the polymer, thereby altering the conformation of the polypeptide. The alteration of the conformation of the polypeptide may increase or decrease the binding affinity of the polypeptide for a substrate bound by the polypeptide, or alter the catalytic rate of an enzyme. Typically, the polymer is a polynucleotide or polypeptide.
Claims17
A method of altering the conformation of a polypeptide, wherein the polypeptide is a sugar-binding protein or a kinase, the method comprising:
- (a) covalently attaching a first end of a synthetic single stranded polynucleotide to a first portion of the polypeptide;
- (b) covalently attaching a second end of the single stranded polynucleotide to a second portion of the polypeptide;
- and
- (c) exerting a force of about 1 to about 10 pN on the polypeptide by hybridizing the single stranded polynucleotide to a complementary polynucleotide to alter the mechanical tension on the polypeptide, wherein the alteration of mechanical tension on the polypeptide effects an alteration of (i) the binding affinity for a kinase substrate, (ii) the binding affinity for a sugar, or (iii) the catalytic rate of the kinase, thereby altering the conformation of the polypeptide.
A composition comprising a polypeptide coupled to a synthetic single stranded polynucleotide, wherein the polypeptide is a sugar-binding protein or a kinase, wherein a first end of the polynucleotide is covalently attached to a first portion of the polypeptide and a second end of the polynucleotide is covalently attached to a second portion of the polypeptide, and wherein the single stranded polynucleotide is selected so that:
- (a) upon hybridization with a complementary polynucleotide, the polynucleotide coupled to the polypeptide exerts an alteration of mechanical tension on the polypeptide of about 1 to about 10 pN;
- and
- (b) the alteration of mechanical tension on the polypeptide effects an alteration of (i) the binding affinity for a kinase substrate, (ii) the binding affinity for a sugar, or (iii) the catalytic rate of the kinase.
Dependencies are inferred from the claim text.