Two novel rat liver membrane proteins that bind advanced glycosylation endproducts

Relationship to macrophage receptor for glucose-modified proteins

Zhi Yang, Zenji Makita, Yasuhiro Horii, Sharon Brunelle, Anthony Cerami, Prabodh Sehajpal, Manikkam Suthanthiran, Helen Vlassara

Research output: Contribution to journalArticle

149 Citations (Scopus)

Abstract

Advanced glycosylation endproducts (AGEs), the glucose-derived adducts that form nonenzymatically and accumulate on tissue proteins, are implicated in many chronic complications associated with diabetes and aging. We have previously described a monocyte/macrophage surface receptor system thought to coordinate AGE protein removal and tissue remodeling, and purified a corresponding 90-kD AGE-binding protein from the murine RAW 264.7 cell line. To identify AGEbinding proteins in normal animals, the tissue distribution of 125I-AGE rat serum albumin taken up from the blood was determined in rats in vivo. These uptake studies demonstrated that the liver was a major site of AGE protein sequestration. Using a solid-phase assay system involving the immobilization of solubilized membrane proteins onto nitrocellulose to monitor binding activity, and several purification steps including affinity chromatography over an AGE bovine serum albumin matrix, two rat liver membrane proteins were isolated that specifically bound AGES, one migrating at 60 kD (p60) and the other at 90 kD (p90) on SDS-PAGE. NH2-terminal sequence analysis revealed no significant homology between these two proteins nor to any molecules available in sequence databases. Flow cytometric analyses using avian antibodies to purified rat p60 and p90demonstrated that both proteins are present on rat monocytes and macrophages. Competition studies revealed no crossreactivity between the two antisera; anti-p60 and anti-p90 antisera prevented AGE-protein binding to rat macrophages when added alone or in combination. These results indicate that rat liver contains at least two novel and distinct proteins that recognize AGE-modified macromolecules, although p90 may berelated to the previously described 90-kD AGE receptor isolated from RAW 264.7 cells. The constitutive expression ofAGE-binding proteins on rat monocytes and macrophages, and the sequestration ofcirculating AGE-modified proteins by the liver, provides further evidence in support of a role for these molecules in the normal removal of proteins markedas senescent by accumulated glucose-derived covalent addition products, or AGES.

Original languageEnglish
Pages (from-to)515-524
Number of pages10
JournalJournal of Experimental Medicine
Volume174
Issue number3
DOIs
Publication statusPublished - 1 Sep 1991
Externally publishedYes

Fingerprint

Glycosylation
Membrane Proteins
Macrophages
Liver
Proteins
Monocytes
Immune Sera
Carrier Proteins
glucose receptor
Glucose
Collodion
Tissue Distribution
Bovine Serum Albumin
Affinity Chromatography
Protein Binding
Serum Albumin
Immobilization
Sequence Analysis
Polyacrylamide Gel Electrophoresis
Databases

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Two novel rat liver membrane proteins that bind advanced glycosylation endproducts : Relationship to macrophage receptor for glucose-modified proteins. / Yang, Zhi; Makita, Zenji; Horii, Yasuhiro; Brunelle, Sharon; Cerami, Anthony; Sehajpal, Prabodh; Suthanthiran, Manikkam; Vlassara, Helen.

In: Journal of Experimental Medicine, Vol. 174, No. 3, 01.09.1991, p. 515-524.

Research output: Contribution to journalArticle

Yang, Zhi ; Makita, Zenji ; Horii, Yasuhiro ; Brunelle, Sharon ; Cerami, Anthony ; Sehajpal, Prabodh ; Suthanthiran, Manikkam ; Vlassara, Helen. / Two novel rat liver membrane proteins that bind advanced glycosylation endproducts : Relationship to macrophage receptor for glucose-modified proteins. In: Journal of Experimental Medicine. 1991 ; Vol. 174, No. 3. pp. 515-524.
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