Crystal structure of hepatocyte growth factor activator complexed with Kunitz domain inhibitor
AssigneeGenentech, Inc.
InventorsCharles W. Eigenbrot, Jr., Daniel K. Kirchhofer
Life of this patent
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Abstract
The present disclosure provides crystals of and structural coordinates of Hepatocyte growth factor activator with and without bound pseudo-substrate or inhibitor. In a specific embodiment, a crystal structure of activated HGFA complexed with a Kunitz domain inhibitor is provided. The crystals and crystal structures are useful, for example, in the design and synthesis of inhibitors of HGFA.
Claims14
A crystal comprising a polypeptide consisting of the amino acid sequence SEQ ID NO:
- 27, wherein the crystal has a space group symmetry of P21 and has a unit cell dimension of a is 52.53 Å, b is 76.43 Å, and c is 72.15 Å.
A method of producing a crystal comprising:
- a) obtaining a crystallization solution consisting of a 1:1 mixture of a polypeptide solution and a reservoir solution, wherein the polypeptide solution consists of:
- a buffer and a polypeptide consisting of the amino acid sequence SEQ ID NO:
- 27, wherein the buffer consists of 10 mM TRIS pH 8, 200 mM NaC1, 0.1% sodium azide, and 10 mM benzamidine in aqueous solution, and wherein the polypeptide is present at a concentration of 8 mg/ml in the buffer, and wherein the reservoir solution consists of 10% PEG 10K and 0.1 M HEPES pH 7.2 in aqueous solution;
- and
- b) incubating the crystallization solution to obtain the crystal.
A crystal comprising a 1:1 complex of a polypeptide consisting of the amino acid sequence SEQ ID NO:
- 27 with a polypeptide consisting of the amino acid sequence SEQ ID NO:
- 28, and wherein the crystal has a space group symmetry of P3121 and has a unit cell dimension of a, b, and c, where a and b are 76.22 Å, and c is 176.24 Å.
A method of producing a crystal comprising:
- a) obtaining a crystallization solution consisting of a 1:1 mixture of a polypeptide complex solution and a reservoir solution, wherein the polypeptide complex solution consists of:
- a buffer and a 1:1 complex of a polypeptide consisting of the amino acid sequence SEQ ID NO:
- 27 with a polypeptide consisting of the amino acid sequence SEQ ID NO:
- 28, wherein the buffer consists of 10 mM HEPES pH 7.2, 150 mM NaC1, 0.5 mM [3-mercaptoethanol in aqueous solution, and wherein the polypeptide is present at a concentration of 7.6 mg/ml in the buffer, and wherein the reservoir solution consists of 50% MPD, 0.1 M TRIS pH 8.5, and 0.2 M ammonium sulfate in aqueous solution;
- and
- b) incubating the crystallization solution to obtain the crystal.
Dependencies are inferred from the claim text.