US2004157298A1PendingUtilityA1

Modified cytokine receptor protein

52
Priority: Feb 18, 1997Filed: Mar 23, 2004Published: Aug 12, 2004
Est. expiryFeb 18, 2017(expired)· nominal 20-yr term from priority
Inventors:Michael SundstromGunnar Norstedt
A61K 38/00C07K 1/306C07K 14/72C07K 14/715C07K 14/71
52
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

A modified extracellular domain of a cytokine receptor protein is capable of being crystallized without being complexed to a ligand molecule. The receptor preferably is a homo- or heterodimeric receptor having at least one molecule segment which contributes to a disordered structure deleted. A preferred receptor is human growth hormone receptor (hGHR).

Claims

exact text as granted — not AI-modified
1 . A modified extracellular domain of a cytokine receptor protein, capable of being crystallized without being complexed to a ligand molecule.  
     
     
         2 . A modified protein according to  claim 1  being a homo- or heterodimeric cytokine receptor.  
     
     
         3 . A modified protein according to claims  1  or  2  wherein at least one molecule segment which contributes to a disordered structure is deleted.  
     
     
         4 . A modified protein according to  claim 3  truncated in at least one terminal end.  
     
     
         5 . A modified protein according to  claim 4  truncated in its C-terminal end and in its N-terminal end.  
     
     
         6 . A modified protein according to  claim 5  being human growth hormone receptor (hGHR).  
     
     
         7 . A modified human growth hormone receptor (hGHR) according to  claim 6  having 31 or 33 amino acid residues removed in its N-terminal end.  
     
     
         8 . A modified human growth hormone receptor (hGHR) according to  claim 6  or  7  having 3 or 4 amino acid residues removed in its C-terminal end.  
     
     
         9 . A modified human growth hormone receptor (hGHR) according to any of  claims 6  to  8  consisting of residues 32-237, 32-234 or 34-233 of the native molecule.  
     
     
         10 . A modified human growth hormone receptor (hGHR) according to  claim 9  consisting residues 32-237 of the native molecule.  
     
     
         11 . Crystals of a receptor protein according to any of  claims 1  to  10  to any of claims  1 - 10  suitable for binding studies with ligand candidates.  
     
     
         12 . Crystals according to  claim 11 , wherein the contact surface between two molecules is between 200 to 1800 Å 2  (square {dot over (a)}ngström) and more preferably between 100 to 900 Å 2  (square {dot over (a)}ngström).  
     
     
         13 . Crystals according to  claim 11  or  12  containing at least 50% (v/v) of a solvent acceptable for binding studies.  
     
     
         14 . Crystals according to  claim 13  containing about 60 to 80 (v/v) of a solvent.  
     
     
         15 . Crystals according to any of  claims 11  to  14  capable of being frozen with gaseous or liquid nitrogen with maintained capacity of diffraction to at least 3.5 Å by using synchrotron radiation source.  
     
     
         16 . Crystals according to  claim 15  capable of being frozen with gaseous or liquid nitrogen with maintained capacity of diffraction to at least 3.5 Å by using synchrotron radiation source.  
     
     
         17 . Crystals according to any of  claims 11  to  16  capable of being resistant to an addition of up to 10% (v/v) of DMSO (dimethylsulfoxide) and up to 5% (v/v) of DMF (dimethylfluoride) for at least 24 hours.  
     
     
         18 . Crystals according to any of  claims 11  to  17  characterized in that they are formed at pH between 5.0 to 8.5.  
     
     
         19 . Crystals according to  claim 18  characterized in that they are formed at a pH between 7.0 and 8.0.  
     
     
         20 . Crystals according to any of  claims 11  to  17  formed in the presence of one or more salts having a concentration between 0.15 M and 1.0 M.  
     
     
         21 . Crystals according to  claim 20 , wherein the salt(s) is(are) selected from a group consisting of ammonium sulfate, lithium sulfate, sodium phosphate, potassium phosphate, sodium chloride, lithium chloride, ammonium acetate, sodium acetate, magnesium chloride, sodium formate and sodium citrate.  
     
     
         22 . A method of designing drugs with cytokine receptor activity by employing the crystals according to any of  claims 11  to  21  in binding studies with selected ligand candidates.  
     
     
         23 . A method according to  claim 22  involving dimerization of the receptor.  
     
     
         24 . A method according to claims  22  or  23 , wherein the crystals are soaked or co-crystallized with a solution comprising the ligands.  
     
     
         25 . A method according to any  claims 22  to  24 , wherein the receptor is a modified growth hormone receptor investigated with ligands having potential growth hormone activity.  
     
     
         26 . A method of obtaining improved cytokine receptor crystals involving the subsequent steps of: 
 (i) solving the receptor three-dimensional structure complexed to a ligand by crystallographic methods,    (ii) identifying regions of the receptor molecule which may contribute to disorder in a crystalline state,    (iii) producing modified receptor molecules without said regions, and    (iv) crystallizing the modified receptor without the presence of a ligand.    
     
     
         27 . A method according to  claim 26  involving the extracellular part of the receptor.  
     
     
         28 . A method according to  claim 26  or  27 , wherein said receptor is human growth hormone receptor.  
     
     
         29 . A method according to  claim 28 , wherein said ligand is human growth hormone.

Join the waitlist — get patent alerts

Track US2004157298A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.