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.
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.
Salazar
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R
Institute of Medical Biochemistry and Molecular Biology, Ernst Moritz Arndt University, Klinikum/Sauerbruch Street, D-17487 Greifswald, Germany.
Brandt
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R
Krantz
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S
D002352:Carrier Proteins / Q000494:pharmacology; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation; D017127:Glycation End Products, Advanced; D006031:Glycosylation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009000:Monocytes / Q000201:enzymology / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism*; D051744:Protein Kinase C-epsilon; D012333:RNA, Messenger / Q000096:biosynthesis; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D015398:Signal Transduction; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D000093423:Glycated Serum Albumin; D000097796:Nucleolin
D002352:Carrier Proteins; D016207:Cytokines; D017127:Glycation End Products, Advanced; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes; D008565:Membrane Proteins; D018995:Myosin Heavy Chains; D016328:NF-kappa B; D010750:Phosphoproteins; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012709:Serum Albumin; D018808:Transcription Factor AP-1; D000093423:Glycated Serum Albumin; D000097796:Nucleolin; C015482:protein kinase modulator; C497255:PRKCE protein, human