Gardberg, Anna and Dice, Lezlee and Pridgen, Kathleen and Ko, Jan and Patterson, Paul and Ou, Susan and Wetzel, Ronald and Dealwis, Chris (2009) Structures of Aβ-Related Peptide−Monoclonal Antibody Complexes. Biochemistry, 48 (23). pp. 5210-5217. ISSN 0006-2960 http://resolver.caltech.edu/CaltechAUTHORS:20090824-132624445
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Passive immunotherapy (PI) is being explored as a potential therapeutic against Alzheimer’s disease. The most promising antibodies (Abs) used in PI target the EFRH motif of the Aβ N-terminus. The monoclonal anti-Aβ Ab PFA1 recognizes the EFRH epitope of Aβ. PFA1 has a high affinity for Aβ fibrils and protofibrils (0.1 nM), as well as good affinity for Aβ monomers (20 nM). However, PFA1 binds the toxic N-terminally modified pyroglutamate peptide pyro-Glu3-Aβ with a 77-fold loss in affinity compared to the WT Aβ(1−8). Furthermore, our earlier work illustrated PFA1’s potential for cross-reactivity. The receptor tyrosine kinase Ror2, which plays a role in skeletal and bone formation, possesses the EFRH sequence. PFA1 Fab binds the Ror2(518−525) peptide sequence REEFRHEA with a 3-fold enhancement over WT Aβ(1−8). In this work, the crystal structures of the hybridoma-derived PFA1 Fab in complex with pyro-Glu3-Aβ peptide and with a cross-reacting peptide from Ror2 have been determined at resolutions of 1.95 and 2.7 Å, respectively. As with wild-type Aβ, these peptides bind to the Fab via a combination of charge- and shape-complementarity, hydrogen-bonding, and hydrophobic interactions. Comparison of the structures of the four peptides Aβ(1−8), Grip1, pyro-Glu3-Aβ(3−8), and Ror2 in complex with PFA1 shows that the greatest conformational flexibility occurs at residues 2 to 3 and 8 of the peptide. These structures provide a molecular basis of the specificity tolerance of PFA1 and its ability to recognize Aβ N-terminal heterogeneity. The structures provide clues to improving mAb specificity and affinity for pyroglutamate Aβ.
|Additional Information:||© 2009 American Chemical Society. Received January 26, 2009; Revised Manuscript Received April 17, 2009. Publication Date (Web): April 22, 2009. We thank the members of the BMC beamline at BioCARS at APS for assistance with data collection. We also thank Dr. Brad Bennett and Jay Prendergast for proofreading the manuscript. Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Science, under Contract W-31-109-Eng-38. Use of the BioCARS Sector 14 was supported by the National Institutes of Health, National Center for Research Resources, under Grant RR007707. Supporting Information: One table containing hydrogen-bonding data for the PFA1-pyro-Glu3-Aβ and PFA1-Ror2(518−525) structures. This material is available free of charge via the Internet at http://pubs.acs.org.|
|Official Citation:||Structures of Aβ-Related Peptide−Monoclonal Antibody Complexes, Anna Gardberg, Lezlee Dice, Kathleen Pridgen, Jan Ko, Paul Patterson, Susan Ou, Ronald Wetzel, Chris Dealwis Biochemistry 2009 48 (23), 5210-5217|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Ruth Sustaita|
|Deposited On:||08 Sep 2009 21:55|
|Last Modified:||26 Dec 2012 11:14|
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