NMR solution structure of the receptor binding domain of human alpha(2)-macroglobulin.
Human alpha(2)-macroglobulin-proteinase complexes bind to their receptor, the low density lipoprotein receptor-related protein (LRP), through a discrete 138-residue C-terminal receptor binding domain (RBD), which also binds to the beta-amyloid peptide. We have used NMR spectroscopy on recombinantly expressed uniformly (13)C/(15)N-labeled human RBD to determine its three-dimensional structure in solution. Human RBD is a sandwich of two antiparallel beta-sheets, one four-strand and one five-strand, and also contains one alpha-helix of 2.5 turns and an additional 1-turn helical region. The principal alpha-helix contains two lysine residues on the outer face that are known to be essential for receptor binding. A calcium binding site (K(d) approximately 11 mM) is present in the loop region at one end of the beta-sandwich. Calcium binding principally affects this loop region and does not significantly perturb the stable core structure of the domain. The structure and NMR assignments will enable us to examine in solution specific binding of RBD to domains of the receptor and to beta-amyloid peptide.
Huang
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W
Department of Biochemistry and Molecular Biology, College of Medicine, University of Illinois at Chicago, Chicago, Illinois 60612-4316, USA.
Dolmer
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K
Liao
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X
Gettins
P G
PG
D000595:Amino Acid Sequence; D000818:Animals; D002247:Carbon Isotopes; D002417:Cattle; D006801:Humans; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D011971:Receptors, Immunologic / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012996:Solutions; D000511:alpha-Macroglobulins / Q000737:chemistry* / Q000378:metabolism*
D002247:Carbon Isotopes; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D009587:Nitrogen Isotopes; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D012996:Solutions; D000511:alpha-Macroglobulins