The Extracellular Region of the Receptor for Advanced Glycation End Products Is Composed of Two Independent Structural Units

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• 作者：Brian M. Dattilo ; Gü ; nter Fritz ; Estelle Leclerc ; Craig W. Vander Kooi ; Claus W. Heizmann ; Walter J. Chazin
• 刊名：Biochemistry
• 出版年：2007
• 出版时间：June 12, 2007
• 年：2007
• 卷：46
• 期：23
• 页码：6957 - 6970
• 全文大小：2730K
• 年卷期：v.46,no.23(June 12, 2007)
• ISSN：1520-4995

文摘

The receptor for advanced glycation end products (RAGE) is an important cell surface receptorbeing pursued as a therapeutic target because it has been implicated in complications arising from diabetesand chronic inflammatory conditions. RAGE is a single membrane spanning receptor containing a verysmall ~40 residue cytosolic domain and a large extracellular region composed of 3 Ig-like domains. Inthis study, high level bacterial expression systems and purification protocols were generated for theextracellular region of RAGE (sRAGE) and the five permutations of single and tandem domain constructsto enable biophysical and structural characterization of its tertiary and quaternary structure. The structureand stability of each of these six protein constructs was assayed by biochemical methods including limitedproteolysis, dynamic light scattering, CD, and NMR. A homology model of sRAGE was constructed toaid in the interpretation of the experimental data. Our results show that the V and C1 domains are notindependent domains, but rather form an integrated structural unit. In contrast, C2 is attached to VC1 bya flexible linker and is fully independent. The interaction with a known RAGE ligand, Ca²⁺-S100B, wasmapped to VC1, with the major contribution from the V domain but clearly defined secondary effectsfrom the C1 domain. The implications of these results are discussed with respect to models for RAGEsignaling.