Three-dimensional structure of bactericidal/permeability-increasing protein (BPI)
AssigneesXoma Corporation, The Regents of the University of California
InventorsLesa J. Beamer, Stephen F. Carroll, David Eisenberg
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Abstract
The present invention provides a crystallized Bactericidal Permeability-Increasing (BPI) protein; methods for x-ray diffraction analysis to provide x-ray diffraction patterns of sufficiently high resolution for three-dimensional structure determination of the protein, as well as methods for rational drug design, based on using amino acid sequence data and/or x-ray crystallography data provided on computer readable media, as analyzed on a computer system having suitable computer algorithms; and atomic coordinates are provided yielding structural information on the lipid binding and lipid transport protein family that includes BPI, LBP, CETP and PLTP.
Claims6
A method for identifying a mutant of a bactericidal/permeability-increasing (BPI) protein with modified activity, wherein the activity is selected from the group consisting of antibacterial, heparin-binding and endotoxin-binding, said method comprising:
- a. constructing a three-dimensional structure of the BPI protein defined by the atomic coordinates shown in FIG. 6;
- b. employing said three-dimensional structure and modeling methods to identify a mutation site that contributes to said activity of said mutant;
- c. producing said mutant;
- d. assaying said mutant to determine said modified activity of said mutant, wherein the activity is selected from the group consisting of antibacterial, heparin-binding, and endotoxin-binding.
A method for identifying a mutant of bactericidal/permeability-increasing (BPI) protein with modified endotoxin-binding or heparin-binding activity, said method comprising:
- a. constructing a three-dimensional structure of the BPI protein defined by the atomic coordinates shown in FIG. 6;
- b. employing said three-dimensional structure and modeling methods to identify a mutation site that contributes to said activity of said mutant, and wherein said mutation site is selected from one or more amino acid residues of a binding pocket defined in column 1 of FIG. 8 or selected from positions about 17 to about 45, positions about 65 to about 99, or positions about 142 to about 169 of BPI of SEQ ID NO:
- 2;
- c. producing said mutant;
- d. contacting said mutant with said endotoxin or heparin to determine said modified binding activity.
Dependencies are inferred from the claim text.