Modified Phosphatases

Abstract

The invention relates to phosphatases and more in specific to (genetically) modified phosphatases, pharmaceutical compositions comprising (genetically) modified phosphatases and the use of (genetically) modified phosphatases for treating or curing for example sepsis, inflammatory bowel disease or other inflammatory diseases, or renal failure. The invention further relates to a method for producing phosphatases.

1 - 35 . (canceled) 
     
     
         36 . An alkaline phosphatase, wherein the alkaline phosphatase is isolated or recombinant and comprises: a crown domain, and a catalytic domain, wherein the crown domain and the catalytic domain are obtained from different alkaline phosphatases. 
     
     
         37 . The alkaline phosphatase of  claim 36 , wherein the crown domain is the crown domain of ALPP and the catalytic domain is the catalytic domain of ALPI. 
     
     
         38 . The alkaline phosphatase of  claim 36 , wherein the crown domain is the crown domain of ALPI and the catalytic domain is the catalytic domain of ALPP. 
     
     
         39 . The alkaline phosphatase of  claim 36 , further comprising a mutation or a deletion of the amino acid sequence encompassing the consensus glycosylphosphatidylinositol (“GPI”) signal sequence in the GPI signal sequence, further wherein the modification or deletion results in a secreted phosphatase. 
     
     
         40 . An isolated or recombinant phosphatase comprising a mutation in the vicinity of a catalytic residue and/or in a metal ion-coordinating phosphate-binding pocket, wherein the mutation is a mutation as depicted in Table 4, 5, or 6. 
     
     
         41 . The phosphatase of  claim 40 , further comprising a modification in the glycosylphosphatidylinositol (“GPI”) signal sequence wherein the modification results in a non-GPI anchored phosphatase and/or comprising a crown domain and a catalytic domain from different phosphatases. 
     
     
         42 . A nucleic acid sequence encoding the alkaline phosphatase of  claim 36 . 
     
     
         43 . A vector comprising the nucleic acid of  claim 42 . 
     
     
         44 . A host cell comprising the nucleic acid of  claim 42 . 
     
     
         45 . A nucleic acid sequence encoding the phosphatase of  claim 39 . 
     
     
         46 . A vector comprising the nucleic acid of  claim 45 . 
     
     
         47 . A host cell comprising the nucleic acid of  claim 45 . 
     
     
         48 . A nucleic acid sequence encoding the phosphatase of  claim 40 . 
     
     
         49 . A vector comprising the nucleic acid of  claim 48 . 
     
     
         50 . A host cell comprising the nucleic acid of  claim 48 . 
     
     
         51 . A method for producing the phosphatase of  claim 36 , the method comprising: culturing a host cell able to express the phosphatase in a medium comprising Zn 2+  and allowing the cell to produce the phosphatase. 
     
     
         52 . The method according to  claim 51 , further comprising isolating the phosphatase. 
     
     
         53 . A phosphatase produced by the method according to  claim 51 . 
     
     
         54 . A pharmaceutical composition comprising the phosphatase of  claim 36 . 
     
     
         55 . A pharmaceutical composition comprising the phosphatase of  claim 53 . 
     
     
         56 . A method for treating a subject suffering from sepsis, inflammatory bowel disease, inflammation of the epithelial lining of the gut, other inflammatory disease, reduced renal function, and/or renal failure, the method comprising: administering to the subject an effective amount of the alkaline phosphatase of  claim 36 . 
     
     
         57 . A method for dephosphorylating a substrate in an environment having a Zn 2+  concentration lower than 10 μM, the method comprising: utilizing the alkaline phosphatase of  claim 38  to dephosphorylate the substrate. 
     
     
         58 . A method for treating a subject suffering from or believed to be suffering from reduced renal function, renal failure and/or a disease which is accompanied by Zn 2+  deficiency, the method comprising: administering an effective amount of the alkaline phosphatase of  claim 37  to the subject. 
     
     
         59 . The method according to  claim 58 , wherein the disease comprises an inflammatory disease and/or is accompanied by intestinal integrity damage. 
     
     
         60 . The method according to  claim 59 , wherein the disease is selected from the group consisting of autoimmune diseases, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, atherosclerosis, inflammatory bowel disease, inflammation of the epithelial lining of the gut, sepsis, neurodermitis, osteitis, and diseases identified in Table 10. 
     
     
         61 . The alkaline phosphatase of  claim 36 , wherein at least one of the different alkaline phosphatases is a human phosphatase. 
     
     
         62 . The isolated or recombinant phosphatase of  claim 40 , wherein the isolated or recombinant phosphatase is a human phosphatase and/or an alkaline phosphatase. 
     
     
         63 . The method according to  claim 51 , wherein the host cell is a mammalian cell. 
     
     
         64 . The method according to  claim 56 , wherein the subject is a human. 
     
     
         65 . The method according to  claim 57 , wherein the substrate is an adenosine phosphate. 
     
     
         66 . The method according to  claim 58 , wherein the subject is a human.