The endothelial cell binding site for advanced glycation end products consists of a complex

An integral membrane protein and a lactoferrin-like polypeptide

Ann Marie Schmidt, Rozalia Mora, Rong Cao, Shi Du Yan, Jerold Brett, Rajasekhar Ramakrishnan, T. Christopher Tsang, Maya Simionescu, David Stern

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

Advanced glycation end products (AGEs), formed as the result of the extended interaction of proteins with ketoses, modulate central properties of endothelial cells and mononuclear phagocytes by interacting with a cell surface binding site comprised of a novel integral membrane protein (receptor for AGE = RAGE) and a lactoferrin-like polypeptide (LF-L), the latter having sequence identity to lactoferrin (LF). To further understand this cellular binding site, the interaction of RAGE with LF-L and LF was characterized. By ligand blotting and a solid state competitive binding assay, 125 I-LF-L and 125 I-LF bound to RAGE immobilized on nitrocellulose membranes or polypropylene tubes in a time-dependent and reversible manner, demonstrating a high affinity component with K(d) ≃ 100 pM. The interaction of 125 I- LF-L and 125 I-LF with RAGE was independent of iron in LF and was competed by addition of an excess of unlabeled carboxyl-terminal portion of LF. Cross- linking studies with purified 125 I-LF-L and RAGE, in the presence of disuccinimidyl suberate, showed a new, slowly migrating band, corresponding to a complex of RAGE and LF-L, and cross-linking on mouse aortic endothelial cells showed two new slowly migrating bands on immunoblotting visualized with both anti-RAGE IgG and anti-LF-L IgG. These data lead us to propose that the endothelial cell surface binding site for AGEs consists of LF-L bound noncovalently to RAGE anchored in the cell membrane.

Original languageEnglish (US)
Pages (from-to)9882-9888
Number of pages7
JournalJournal of Biological Chemistry
Volume269
Issue number13
StatePublished - Apr 1 1994
Externally publishedYes

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Advanced Glycosylation End Products
Lactoferrin
Endothelial cells
Membrane Proteins
Endothelial Cells
Binding Sites
Peptides
Ketoses
Competitive Binding
Collodion
Polypropylenes
Cell membranes
Phagocytes
Immunoblotting
Crosslinking

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The endothelial cell binding site for advanced glycation end products consists of a complex : An integral membrane protein and a lactoferrin-like polypeptide. / Schmidt, Ann Marie; Mora, Rozalia; Cao, Rong; Yan, Shi Du; Brett, Jerold; Ramakrishnan, Rajasekhar; Tsang, T. Christopher; Simionescu, Maya; Stern, David.

In: Journal of Biological Chemistry, Vol. 269, No. 13, 01.04.1994, p. 9882-9888.

Research output: Contribution to journalArticle

Schmidt, AM, Mora, R, Cao, R, Yan, SD, Brett, J, Ramakrishnan, R, Tsang, TC, Simionescu, M & Stern, D 1994, 'The endothelial cell binding site for advanced glycation end products consists of a complex: An integral membrane protein and a lactoferrin-like polypeptide', Journal of Biological Chemistry, vol. 269, no. 13, pp. 9882-9888.
Schmidt, Ann Marie ; Mora, Rozalia ; Cao, Rong ; Yan, Shi Du ; Brett, Jerold ; Ramakrishnan, Rajasekhar ; Tsang, T. Christopher ; Simionescu, Maya ; Stern, David. / The endothelial cell binding site for advanced glycation end products consists of a complex : An integral membrane protein and a lactoferrin-like polypeptide. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 13. pp. 9882-9888.
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