Materials and methods related to sodium/potassium adenosine triphosphase and cholesterol
This invention is based in part on the elucidation of new structural conformations and functions of the sodium/potassium adenosine triphosphate synthase (Na/K ATPase), and especially elucidation of new binding sites and interactions. The present invention provides practical applications of several surprising structural and functional relationships between Na/K ATPase and compounds which interact with Na/K ATPase. Disclosure of these structures and relationships provides insight and practical solutions to chemically affecting not only the Na/K ATPase interactions, but also regulators known to be upstream and downstream.
1. A method to affect cholesterol transport in a cell, comprising:
administering to the cell a composition of matter comprising an amino acid compound of 17 residues comprising at least ten consecutive amino acid residues of a cholesterol recognition/interaction amino acid sequence and consensus pattern (CRAC) sequence having SEQ ID NO:2, or conservative substitutions of the at least ten consecutive amino acid residues of SEQ ID NO:2, wherein the compound is capable of binding cholesterol and affecting sodium/potassium adenosine triphosphate synthase (Na/K ATPase) cholesterol-binding activity by antagonizing the CRAC domain on an α1 subunit of Na/K ATPase.
2. The method of claim 1 , wherein Na/K ATPase cholesterol-binding activity is affected in a manner selected from the group consisting of: decreasing; increasing; eliminating; periodically disrupting; and periodically enhancing.
3. A method to ameliorate neurodegeneration due to pathogenic intracellular cholesterol accumulation in a cell in need of such amelioration, comprising: administering to the cell a composition of matter comprising an amino acid compound of 17 residues comprising at least ten consecutive amino acid residues of a cholesterol recognition/interaction amino acid sequence and consensus pattern (CRAC) sequence having SEQ ID NO:2, or conservative substitutions of the at least ten consecutive amino acid residues of SEQ ID NO:2, wherein the compound is capable of binding cholesterol and decreasing the cholesterol binding activity of sodium/potassium adenosine triphosphate synthase (Na/K ATPase) by antagonizing the CRAC domain in an α1 subunit of Na/K ATPase.
4. A method of treating Niemann Pick, type C1 disease, comprising: administering to the cell a composition of matter comprising an amino acid compound of 17 residues comprising at least ten consecutive amino acid residues of a cholesterol recognition/interaction amino acid sequence and consensus pattern (CRAC) sequence having SEQ ID NO:2, or conservative substitutions of the at least ten consecutive amino acid residues of SEQ ID NO:2, wherein the compound is capable of binding cholesterol and decreasing the ability of sodium/potassium adenosine triphosphate synthase (Na/K ATPase) to bind to cholesterol by antagonizing the CRAC domain of an α1 subunit of Na/K ATPase.
5. The method of claim 4 , wherein the decrease is accomplished by inhibiting the CRAC domain of the α1 subunit of Na/K ATPase.
6. A method to identify compositions capable of treating cholesterol-related disease states, comprising
a) contacting a test sample with a composition comprising an amino acid compound of 17 residues comprising at least ten consecutive amino acid residues or conservative substitutions of the at least ten consecutive amino acid residues of a cholesterol recognition/interaction amino acid sequence and consensus pattern (CRAC) domain in an α1 subunit of Na/K ATPase having SEQ ID NO:2; and
b) identifying if step a) results in antagonizing the ability of cholesterol to bind to the CRAC domain of the α1 subunit of Na/K ATPase.
7. A method to identify compositions capable of treating at least one disease state, comprising
a) contacting a test sample with a composition comprising an amino acid compound of 17 residues comprising at least ten consecutive amino acid residues or conservative substitutions of the at least ten consecutive amino acid residues of a cholesterol recognition/interaction amino acid sequence and consensus pattern (CRAC) domain in an α1 subunit of Na/K ATPase having SEQ ID NO:2; and
b) identifying if step a) results in antagonizing the ability of cholesterol to bind to the CRAC domain of the α1 subunit of Na/K ATPase, wherein the disease state is selected from the group consisting of: NPC1; pathogenic lipid accumulation; vascular disease; heart attack; stroke; overweight; obesity; diabetes; metabolic syndrome; thyroid malfunction; medication side effect; atherosclerosis; heart failure; heart disease; Alzheimer's disease; Parkinson disease; Huntington disease; Tay Sachs disease and neurodegenerative disease.
8. A method to affect the trafficking and expression of caveolin-1 in a cell, comprising:
administering to the cell, a composition of matter comprising an amino acid compound of 17 residues comprising at least ten consecutive amino acid residues of a cholesterol recognition/interaction amino acid sequence and consensus pattern (CRAC) sequence having SEQ ID NO:2, or conservative substitutions of the at least ten consecutive amino acid residues of SEQ ID NO:2, wherein the compound is capable of binding cholesterol and down-regulating plasma membrane α1 subunit of sodium/potassium adenosine triphosphate synthase (Na/K ATPase) the Na/K ATPase having the CRAC domain in an α1 subunit of Na/K ATPase.
9. A method to treat Niemann Pick, type C1 disease (NPC1 disease), comprising:
administering to a subject having Niemann Pick, type C1 disease, a composition of matter comprising an amino acid compound of 17 residues comprising at least ten consecutive amino acid residues of a cholesterol recognition/interaction amino acid sequence and consensus pattern (CRAC) sequence having SEQ ID NO:2, or conservative substitutions of the at least ten consecutive amino acid residues of SEQ ID NO:2, wherein the compound is capable of binding cholesterol and altering expression of the α1 subunit of sodium/potassium adenosine triphosphate synthase (Na/K ATPase) the Na/K ATPase having the CRAC domain in an α1 subunit of Na/K ATPase so as to ameliorate the symptoms of NPC1 disease.
10. The method of claim 1 , wherein the cell is a mammal cell.
11. The method of claim 3 , wherein the cell is a mammal cell.
12. The method of claim 8 , wherein the cell is a mammal cell.
13. The method of claim 10 , wherein the mammal cell is selected from the group consisting of: liver cells; kidney cells; brain cells; nerve cells; pancreatic cells; lung cells; skin cells; heart cells; rodent cells; human cells; mouse cells; rat cells; guinea pig cells; dog cells; and, monkey cells.
14. The method of claim 11 , wherein the mammal cell is selected from the group consisting of: liver cells; kidney cells; brain cells; nerve cells; pancreatic cells; lung cells; skin cells; heart cells; rodent cells; human cells; mouse cells; rat cells; guinea pig cells; dog cells; and, monkey cells.
15. The method of claim 12 , wherein the mammal cell is selected from the group consisting of: liver cells; kidney cells; brain cells; nerve cells; pancreatic cells; lung cells; skin cells; heart cells; rodent cells; human cells; mouse cells; rat cells; guinea pig cells; dog cells; and, monkey cells.