Binding of Amadori glucose-modified albumin by the monocytic cell line MonoMac 6 activates protein kinase C epsilon protein tyrosine kinases and the transcription factors AP-1 and NF-kappaB

Glycoconj J. 2001 Oct;18(10):769-77. doi: 10.1023/a:1021151417556.

Abstract

An affinity purification procedure is employed for the isolation of FL-specific binding proteins from MM6 cell membranes using magnetobeads coated with glycated polylysine and elution with FL and glycated 6-aminocaproic acid. Two main binding proteins were identified as membrane-bound nucleolin and cellular myosin heavy chain, which are glycosylated. This study shows that in these cells binding of short-term glycated albumin leads to activation of PKC, especially its isoform epsilon and this is linked to translocation of AP-1 and NF-kappaB into the nucleus. Consequently, an increased formation of IL-1ss mRNA is observed. The PKC inhibitor GO6976 prevents all these effects. Glycated albumin also stimulates activation of PTK. The PTK inhibitor genistein prevents activation of AP-1 indicating that PTK is also involved in this process, whereas NF-kappaB translocation is only dependent on PKC activation.

MeSH terms

  • Carrier Proteins / pharmacology
  • Cell Line
  • Cytokines / biosynthesis
  • Electrophoretic Mobility Shift Assay
  • Enzyme Activation
  • Glycation End Products, Advanced
  • Glycosylation
  • Humans
  • Intracellular Signaling Peptides and Proteins*
  • Isoenzymes / metabolism*
  • Membrane Proteins / chemistry
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism
  • Monocytes / enzymology
  • Monocytes / metabolism*
  • Myosin Heavy Chains / chemistry
  • Myosin Heavy Chains / isolation & purification
  • Myosin Heavy Chains / metabolism
  • NF-kappa B / metabolism*
  • Phosphoproteins / chemistry
  • Phosphoproteins / metabolism
  • Protein Binding
  • Protein Kinase C / metabolism*
  • Protein Kinase C-epsilon
  • RNA, Messenger / biosynthesis
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Serum Albumin / chemistry
  • Serum Albumin / metabolism*
  • Serum Albumin / pharmacology
  • Signal Transduction
  • Transcription Factor AP-1 / metabolism*

Substances

  • Carrier Proteins
  • Cytokines
  • Glycation End Products, Advanced
  • Intracellular Signaling Peptides and Proteins
  • Isoenzymes
  • Membrane Proteins
  • NF-kappa B
  • Phosphoproteins
  • RNA, Messenger
  • RNA-Binding Proteins
  • Serum Albumin
  • Transcription Factor AP-1
  • glycated serum albumin
  • nucleolin
  • protein kinase modulator
  • PRKCE protein, human
  • Protein Kinase C
  • Protein Kinase C-epsilon
  • Myosin Heavy Chains