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Protein Sci. Oct 2000; 9(10): 1889–1897.
PMCID: PMC2144472

Structure of a rat alpha 1-macroglobulin receptor-binding domain dimer.

Abstract

Alpha-macroglobulin inhibits a broad spectrum of proteinases by forming macromolecular cages inside which proteinases are cross-linked and trapped. Upon formation of a complex with proteinase, alpha-macroglobulin undergoes a large conformational change that results in the exposure of its receptor-binding domain (RBD). Engagement of this domain by alpha-macroglobulin receptor permits clearance of the alpha-macroglobulin: proteinase complex from circulation. The crystal structure of rat alpha1-macroglobulin RBD has been determined at 2.3 A resolution. The RBD is composed of a nine-stranded beta-sandwich and a single alpha-helix that has been implicated as part of the receptor binding site and that lies on the surface of the beta-sandwich. The crystallographic asymmetric unit contains a dimer of RBDs related by approximate twofold symmetry such that the putative receptor recognition sites of the two monomers are contiguous. By gel filtration and ultracentrifugation, it is shown that RBD dimers form in solution with a dissociation constant of approximately 50 microM. The structure of the RBD dimer might mimic a conformation of transformed alpha-macroglobulin in which the proposed receptor binding residues are exposed on one face of the dimer. A pair of phenylalanine residues replaces a cystine that is conserved in other members of the macroglobulin family. These residues participate in a network of aromatic side-chain interactions that appears to stabilize the dimer interface.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Andersen GR, Koch TJ, Dolmer K, Sottrup-Jensen L, Nyborg J. Low resolution X-ray structure of human methylamine-treated alpha 2-macroglobulin. J Biol Chem. 1995 Oct 20;270(42):25133–25141. [PubMed]
  • Armstrong PB, Quigley JP. Alpha2-macroglobulin: an evolutionarily conserved arm of the innate immune system. Dev Comp Immunol. 1999 Jun-Jul;23(4-5):375–390. [PubMed]
  • Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE. The Protein Data Bank. Nucleic Acids Res. 2000 Jan 1;28(1):235–242. [PMC free article] [PubMed]
  • Birkenmeier G, Osman AA, Kopperschläger G, Mothes T. Epitope mapping by screening of phage display libraries of a monoclonal antibody directed against the receptor binding domain of human alpha2-macroglobulin. FEBS Lett. 1997 Oct 20;416(2):193–196. [PubMed]
  • Blacker D, Wilcox MA, Laird NM, Rodes L, Horvath SM, Go RC, Perry R, Watson B, Jr, Bassett SS, McInnis MG, et al. Alpha-2 macroglobulin is genetically associated with Alzheimer disease. Nat Genet. 1998 Aug;19(4):357–360. [PubMed]
  • Borth W, Luger TA. Identification of alpha 2-macroglobulin as a cytokine binding plasma protein. Binding of interleukin-1 beta to "F" alpha 2-macroglobulin. J Biol Chem. 1989 Apr 5;264(10):5818–5825. [PubMed]
  • Brünger AT, Adams PD, Clore GM, DeLano WL, Gros P, Grosse-Kunstleve RW, Jiang JS, Kuszewski J, Nilges M, Pannu NS, et al. Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905–921. [PubMed]
  • Burley SK, Petsko GA. Aromatic-aromatic interaction: a mechanism of protein structure stabilization. Science. 1985 Jul 5;229(4708):23–28. [PubMed]
  • Delain E, Barray M, Tapon-Bretaudiere J, Pochon F, Marynen P, Cassiman JJ, Van den Berghe H, Van Leuven F. The molecular organization of human alpha 2-macroglobulin. An immunoelectron microscopic study with monoclonal antibodies. J Biol Chem. 1988 Feb 25;263(6):2981–2989. [PubMed]
  • Dolmer K, Huang W, Gettins PG. NMR solution structure of complement-like repeat CR3 from the low density lipoprotein receptor-related protein. Evidence for specific binding to the receptor binding domain of human alpha(2)-macroglobulin. J Biol Chem. 2000 Feb 4;275(5):3264–3269. [PubMed]
  • Eggertsen G, Hudson G, Shiels B, Reed D, Fey GH. Sequence of rat alpha 1-macroglobulin, a broad-range proteinase inhibitor from the alpha-macroglobulin-complement family. Mol Biol Med. 1991 Apr;8(2):287–302. [PubMed]
  • Fass D, Blacklow S, Kim PS, Berger JM. Molecular basis of familial hypercholesterolaemia from structure of LDL receptor module. Nature. 1997 Aug 14;388(6643):691–693. [PubMed]
  • Gliemann J, Nykjaer A, Petersen CM, Jørgensen KE, Nielsen M, Andreasen PA, Christensen EI, Lookene A, Olivecrona G, Moestrup SK. The multiligand alpha 2-macroglobulin receptor/low density lipoprotein receptor-related protein (alpha 2MR/LRP). Binding and endocytosis of fluid phase and membrane-associated ligands. Ann N Y Acad Sci. 1994 Sep 10;737:20–38. [PubMed]
  • Herz J, Hamann U, Rogne S, Myklebost O, Gausepohl H, Stanley KK. Surface location and high affinity for calcium of a 500-kd liver membrane protein closely related to the LDL-receptor suggest a physiological role as lipoprotein receptor. EMBO J. 1988 Dec 20;7(13):4119–4127. [PMC free article] [PubMed]
  • Holtet TL, Nielsen KL, Etzerodt M, Moestrup SK, Gliemann J, Sottrup-Jensen L, Thøgersen HC. Receptor-binding domain of human alpha 2-macroglobulin. Expression, folding and biochemical characterization of a high-affinity recombinant derivative. FEBS Lett. 1994 May 16;344(2-3):242–246. [PubMed]
  • Huang W, Dolmer K, Gettins PG. NMR solution structure of complement-like repeat CR8 from the low density lipoprotein receptor-related protein. J Biol Chem. 1999 May 14;274(20):14130–14136. [PubMed]
  • Huang W, Dolmer K, Liao X, Gettins PG. NMR solution structure of the receptor binding domain of human alpha(2)-macroglobulin. J Biol Chem. 2000 Jan 14;275(2):1089–1094. [PubMed]
  • Hughes SR, Khorkova O, Goyal S, Knaeblein J, Heroux J, Riedel NG, Sahasrabudhe S. Alpha2-macroglobulin associates with beta-amyloid peptide and prevents fibril formation. Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):3275–3280. [PMC free article] [PubMed]
  • Janin J, Miller S, Chothia C. Surface, subunit interfaces and interior of oligomeric proteins. J Mol Biol. 1988 Nov 5;204(1):155–164. [PubMed]
  • Jenner L, Husted L, Thirup S, Sottrup-Jensen L, Nyborg J. Crystal structure of the receptor-binding domain of alpha 2-macroglobulin. Structure. 1998 May 15;6(5):595–604. [PubMed]
  • Jones S, Thornton JM. Principles of protein-protein interactions. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):13–20. [PMC free article] [PubMed]
  • Moestrup SK, Gliemann J. Analysis of ligand recognition by the purified alpha 2-macroglobulin receptor (low density lipoprotein receptor-related protein). Evidence that high affinity of alpha 2-macroglobulin-proteinase complex is achieved by binding to adjacent receptors. J Biol Chem. 1991 Jul 25;266(21):14011–14017. [PubMed]
  • Moestrup SK, Kaltoft K, Sottrup-Jensen L, Gliemann J. The human alpha 2-macroglobulin receptor contains high affinity calcium binding sites important for receptor conformation and ligand recognition. J Biol Chem. 1990 Jul 25;265(21):12623–12628. [PubMed]
  • Neels JG, van Den Berg BM, Lookene A, Olivecrona G, Pannekoek H, van Zonneveld AJ. The second and fourth cluster of class A cysteine-rich repeats of the low density lipoprotein receptor-related protein share ligand-binding properties. J Biol Chem. 1999 Oct 29;274(44):31305–31311. [PubMed]
  • Nicholls A, Sharp KA, Honig B. Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins. 1991;11(4):281–296. [PubMed]
  • Nielsen KL, Holtet TL, Etzerodt M, Moestrup SK, Gliemann J, Sottrup-Jensen L, Thogersen HC. Identification of residues in alpha-macroglobulins important for binding to the alpha2-macroglobulin receptor/Low density lipoprotein receptor-related protein. J Biol Chem. 1996 May 31;271(22):12909–12912. [PubMed]
  • Nielsen KL, Sottrup-Jensen L, Fey GH, Thøgersen HC. Expression and refolding of a high-affinity receptor binding domain from rat alpha 1-macroglobulin. FEBS Lett. 1995 Oct 16;373(3):296–298. [PubMed]
  • O'Connor-McCourt MD, Wakefield LM. Latent transforming growth factor-beta in serum. A specific complex with alpha 2-macroglobulin. J Biol Chem. 1987 Oct 15;262(29):14090–14099. [PubMed]
  • RAMACHANDRAN GN, RAMAKRISHNAN C, SASISEKHARAN V. Stereochemistry of polypeptide chain configurations. J Mol Biol. 1963 Jul;7:95–99. [PubMed]
  • Sottrup-Jensen L. Alpha-macroglobulins: structure, shape, and mechanism of proteinase complex formation. J Biol Chem. 1989 Jul 15;264(20):11539–11542. [PubMed]
  • Sottrup-Jensen L, Gliemann J, Van Leuven F. Domain structure of human alpha 2-macroglobulin. Characterization of a receptor-binding domain obtained by digestion with papain. FEBS Lett. 1986 Sep 1;205(1):20–24. [PubMed]
  • Stoops JK, Schroeter JP, Kolodziej SJ, Strickland DK. Structure-function relationships of human alpha 2-macroglobulin. Three-dimensional structures of native alpha 2-macroglobulin and its methylamine and chymotrypsin derivatives. Ann N Y Acad Sci. 1994 Sep 10;737:212–228. [PubMed]
  • Swenson RP, Howard JB. Characterization of alkylamine-sensitive site in alpha 2-macroglobulin. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4313–4316. [PMC free article] [PubMed]
  • Wärmegård B, Martin N, Johansson S. cDNA cloning and sequencing of rat alpha 1-macroglobulin. Biochemistry. 1992 Mar 3;31(8):2346–2352. [PubMed]
  • Wilson C, Wardell MR, Weisgraber KH, Mahley RW, Agard DA. Three-dimensional structure of the LDL receptor-binding domain of human apolipoprotein E. Science. 1991 Jun 28;252(5014):1817–1822. [PubMed]
  • Woessner JF., Jr Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J. 1991 May;5(8):2145–2154. [PubMed]

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