Structure of a receptor-binding fragment of reelin and mutational analysis reveal a recognition mechanism similar to endocytic receptors.
Reelin, a large secreted protein implicated in the cortical development of the mammalian brain, is composed of eight tandem concatenations of "reelin repeats" and binds to neuronal receptors belonging to the low-density lipoprotein receptor gene family. We found that both receptor-binding and subsequent Dab1 phosphorylation occur solely in the segment ... spanning the fifth and sixth reelin repeats (R5-6). Monomeric fragment exhibited a suboptimal level of signaling activity and artificial oligomerization resulted in a 10-fold increase in activity, indicating the critical importance of higher-order multimerization in physiological reelin. A 2.0-A crystal structure from the R5-6 fragment revealed not only a unique domain arrangement wherein two repeats were aligned side by side with the same orientation, but also the unexpected presence of bound Zn ions. Structure-guided alanine mutagenesis of R5-6 revealed that two Lys residues (Lys-2360 and Lys-2467) constitute a central binding site for the low-density lipoprotein receptor class A module in the receptor, indicating a strong similarity to the ligand recognition mode shared among the endocytic lipoprotein receptors.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Amino Acid Sequence, Animals, Binding Sites, CHO Cells, Cell Adhesion Molecules, Neuronal, Cell Line, Cells, Cultured, Cerebral Cortex, Clone Cells, Conserved Sequence, Cricetinae, Cricetulus, Crystallography, X-Ray, DNA Mutational Analysis, Disulfides, Electroporation, Extracellular Matrix Proteins, Glutamic Acid, Histidine, Humans, LDL-Receptor Related Proteins, Ligands, Mice, Models, Chemical, Models, Molecular, Molecular Sequence Data, Mutagenesis, Nerve Tissue Proteins, Neurons, Peptide Fragments, Phosphorylation, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Receptors, Lipoprotein, Recombinant Fusion Proteins, Serine Endopeptidases, Zinc
Adaptor Proteins, Signal Transducing, Amino Acid Sequence, Animals, Binding Sites, CHO Cells, Cell Adhesion Molecules, Neuronal, Cell Line, Cells, Cultured, Cerebral Cortex, Clone Cells, Conserved Sequence, Cricetinae, Cricetulus, Crystallography, X-Ray, DNA Mutational Analysis, Disulfides, Electroporation, Extracellular Matrix Proteins, Glutamic Acid, Histidine, Humans, LDL-Receptor Related Proteins, Ligands, Mice, Models, Chemical, Models, Molecular, Molecular Sequence Data, Mutagenesis, Nerve Tissue Proteins, Neurons, Peptide Fragments, Phosphorylation, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Receptors, Lipoprotein, Recombinant Fusion Proteins, Serine Endopeptidases, Zinc
Proc. Natl. Acad. Sci. U.S.A.
Date: Jun. 12, 2007
PubMed ID: 17548821
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