Variant integrin polypeptides and uses thereof

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Chemical modification or the reaction product thereof – e.g.,...

Reexamination Certificate

Rate now

  [ 0.00 ] – not rated yet Voters 0   Comments 0

Details

C435S007100, C530S324000

Reexamination Certificate

active

07323552

ABSTRACT:
Polypeptides comprising all or part of a variant integrin α subunit A domain and its flanking region are described. In solution or in membrane-associated form, the A domain polypeptides of the invention exists predominantly in a high affinity conformation. In the polypeptides of the invention, referred to as variant integrin polypeptides, a crucial isoleucine or glutamic acid residue is altered. For example, the glutamic acid can be either deleted or replaced with different amino acids residue, e.g., glutamine, aspartic acid, or alanine The variant integrin polypeptides of the invention selectively impair binding of activation-dependent ligands, but not independent ligands. They are useful in screening assays for the identification of molecules that enhance binding of variant polypeptides with impaired binding. In addition, they are useful in distinguishing between activation-dependent ligands and activation-independent ligands. They are also useful for generating antibodies, e.g., monoclonal antibodies, which bind to the impaired form of an integrin. Some such antibodies recognize an epitope that is either not present or not accessible on an integrin that is in the high affinity conformation. The variant integrin polypeptides of the invention can be derived from any integrin α subunit that could be used therapeutically.

REFERENCES:
patent: 5985278 (1999-11-01), Mitjans et al.
patent: 2003/0078375 (2003-04-01), Arnaout et al.
patent: WO 00/59878 (2000-10-01), None
patent: WO 00/60355 (2000-10-01), None
patent: WO 02/04521 (2002-01-01), None
patent: WO 02/09737 (2002-02-01), None
patent: WO 02/31511 (2002-04-01), None
Lazar E et al., Transforming growth factor alpha: mutation of aspartic acid 47 and leucine 48 results in different biological activities. Mol Cell Biol. 8:1247-1252, 1988.
Burgess, et al. Possible dissociation of the heparin-binding and mitogenic activities of heparin-binding growth factor-1 from its receptor-binding activities by site-directed mutagenesis of a single lysine residue. J Cell Biol. 111:2129-2138, 1990.
Baldwin et al., “Cation binding to the integrin CD11b I domain and activation model assessment”,Structure, 6:923-935 (1998).
Edwards et al., “Mapping the Intercellular Adhesion Molecule-1 and -2 Binding Site on the Inserted Domain of Leukocyte Function-associated Antigen-1”,The Journal of Biological Chemistry, 273:28937-28944 (1998).
Emsley et al., “Structural Basis of Collagen Recognition by Integrin α2β1”,Cell, 100:47-56 (2000).
Emsley et al., “Crystal Structure of the I Domain from Integrin α2β1”,The Jounal of Biological Chemistry, 272:28512-28517 (1997).
Feng et al., “Peptides Derived from the Complementarity-determining Regions of Anti-Mac-1 Antibodies Block Intercellular Adhesion Molecule-1 Interaction with Mac-1”,The Jounal of Biological Chemistry, 273:5625-5630 (1998).
Kamata T and Takada Y, “Direct Binding of Collagen to the I Domain of Integrin α2β1 (VLA-2, CD49b/CD29) in a Divalent Cation-independent Manner”,The Journal of Biolocal Chemistry, 269:26006-26010 (1994).
Kern et al., “The Role of the I Domain in Ligand Binding of the Human Integrin α1β1”,The Journal of Biological Chemistry, 269:22811-22816 (1994).
Lee et al., “Crystal Structure of the A Domain from the α Subunit of Integrin CR3 (CD11b/CD18)”Cell, 80:631-638 (1995).
Lee et al., “Two conformations of the integrin A-domain (I-domain): a pathway for activation?”,Structure, 3:1333-1340 (1995).
Legge et al., “NMR Solution Structure of the Inserted Domain of Human Leukocyte Function Associated Antigen-1”,Journal of Molecular Biology, 295:1251-1264 (2000).
Li et al., “Two Functional States of the CD11b A-Domain: Correlations with Key Features of Two Mn2+-complexed Crystal Structures”,The Journal of Cell Biology, 143:1523-1534 (1998).
Michishita et al., “A Novel Divalent Cation-Binding Site in the A Domain of the β2 Integrin CR3 (CD11b/CD18) Is Essential for Ligand Binding”,Cell, 72:857-867 (1993).
Nolte et al., “Crystal structure of the α1β1 integrin I-domain: insights into integrin I-domain function”,FEBS Letters, 452:379-385 (1999).
Oxvig et al., “Conformational changes in tertiary structure near the ligand binding site of an integrin I domain”,Proc. Natl. Acad. Sci. USA, 96:2215-2220 (1999).
Rieu et al., “Solvent-accessible Residues on the Metal Ion-dependent Adhesion Site Face of Integrin CR3 Mediate Its Binding to the Neutrophil Inhibitory Factor”,The Journal of Biological Chemistry, 271:15858-15861 (1996).
Smith JW and Cheresh DA, “The Arg-Cly-Asp Binding Domain of the Vitronectin Receptor”,The Journal of Biological Chemistry, 263:18726-18731 (1988).
Xiong et al., “An Isoleucine-based Allosteric Switch Controls Affinity . . . ” J. of Biol. Chem. 275:38762-38767, 2000.
Zhang L and Plow EF, “A Discrete Site Modulates Activation of I Domains”,The Journal of Biological Chemistry, 271:29953-29957 (1996).
Zhang L and Plow EF, “Amino Acid Sequences within the αSubunit of Integrin αMβ2(Mac-1) Critical for Specific Recognition of C3bi”,Biochemistry, 38:8064-8071 (1999).
Xiong et al., “An Isoleucine-based Allosteric Switch Controls Affinity . . . ”J. of Biol. Chem. 275:38762-38767, 2000.
Huth et al., NMR and Mutagenesis Evidence for and I Domain Allosteric Site . . . : Proc. Nat;l. Acad. Sci. 97(10):5235, 2000.
Burgess et al. “Possible dissociation of the Heparin-Binding and Mitogenic Activities”, J. Cell. Biol. III: 2129-2138 (1990).
Lazar et al. “Transforming Growth Factor Alpha . . . ” Mol. Cell. Biol. 8: 1247-1252 (1988).

LandOfFree

Say what you really think

Search LandOfFree.com for the USA inventors and patents. Rate them and share your experience with other people.

Rating

Variant integrin polypeptides and uses thereof does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Variant integrin polypeptides and uses thereof, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Variant integrin polypeptides and uses thereof will most certainly appreciate the feedback.

Rate now

     

Profile ID: LFUS-PAI-O-2794607

  Search
All data on this website is collected from public sources. Our data reflects the most accurate information available at the time of publication.