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AssigneeAstraZenca AB
InventorsPhilippe Cronet, Wolfgang Knecht, Cecilia Ann-Christin Malmborg Hager, Mats Andersson, Christina Furebring
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This invention relates to mutant forms of carboxypeptidase U with increased thermal stability relative to wild-type. In addition to the individual thermal stabilizing mutations identified (at positions 166, 204, 219, 230, 251, 315), the inventors have identified a region (S327-H357) that is crucial to the stability of CPU. The invention relates to nucleic acid encoding such mutant forms and the polypeptides encoded thereby. The invention also relates to methods and materials for making CPU mutants with increased thermal stability relative to wild-type and their use, for example to produce crystals of CPU or proCPU for 3-dimensional structure determination, or in therapy.
A carboxypeptidase U (CPU) mutant polypeptide having greater thermal stability than the wild-type polypeptides, which mutant possesses at least two amino acid substitutions relative to the wild-type polypeptide, at least one of which is located at an amino acid residue position relative to SEQ ID NO:
A method of causing blood to clot comprising contacting the blood with an effective amount of a CPU mutant comprising the amino acid sequence according to SEQ ID NO:
The method of producing wild-type CPU or proCPU crystals, comprising mixing together purified CPU or proCPU polypeptide with a Fab fragment directed to all or part of amino acids from positions 327 to 357 inclusive (according to the position in SEQ ID NO:
Dependencies are inferred from the claim text.