High glucose and insulin promote O-GlcNAc modification of proteins, including alpha-tubulin.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Author(s): Walgren JL;Walgren JL; Vincent TS; Schey KL; Buse MG
  • Source:
    American journal of physiology. Endocrinology and metabolism [Am J Physiol Endocrinol Metab] 2003 Feb; Vol. 284 (2), pp. E424-34. Date of Electronic Publication: 2002 Oct 22.
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: American Physiological Society Country of Publication: United States NLM ID: 100901226 Publication Model: Print-Electronic Cited Medium: Print ISSN: 0193-1849 (Print) Linking ISSN: 01931849 NLM ISO Abbreviation: Am J Physiol Endocrinol Metab Subsets: MEDLINE
    • Publication Information:
      Original Publication: Bethesda, MD. : American Physiological Society
    • Subject Terms:
      Glucose/*pharmacology ; Hypoglycemic Agents/*pharmacology ; Insulin/*pharmacology ; Muscle Cells/*metabolism ; Tubulin/*metabolism; Acetylglucosamine/metabolism ; Animals ; Cells, Cultured ; Glycosylation/drug effects ; HSP70 Heat-Shock Proteins/metabolism ; Insulin Resistance/physiology ; Mass Spectrometry ; Membrane Proteins/metabolism ; Muscle Cells/cytology ; Sp1 Transcription Factor/metabolism
    • Abstract:
      Increased flux through the hexosamine biosynthesis pathway has been implicated in the development of glucose-induced insulin resistance and may promote the modification of certain proteins with O-linked N-acetylglucosamine (O-GlcNAc). L6 myotubes (a model of skeletal muscle) were incubated for 18 h in 5 or 25 mM glucose with or without 10 nM insulin. As assessed by immunoblotting with an O-GlcNAc-specific antibody, high glucose and/or insulin enhanced O-GlcNAcylation of numerous proteins, including the transcription factor Sp1, a known substrate for this modification. To identify novel proteins that may be O-GlcNAc modified in a glucose concentration/insulin-responsive manner, total cell membranes were separated by one- or two-dimensional gel electrophoresis. Selected O-GlcNAcylated proteins were identified by mass spectrometry (MS) analysis. MS sequencing of tryptic peptides identified member(s) of the heat shock protein 70 (HSP70) family and rat alpha-tubulin. Immunoprecipitation/immunoblot studies demonstrated several HSP70 isoforms and/or posttranslational modifications, some with selectively enhanced O-GlcNAcylation following exposure to high glucose plus insulin. In conclusion, in L6 myotubes, Sp1, membrane-associated HSP70, and alpha-tubulin are O-GlcNAcylated; the modification is markedly enhanced by sustained increased glucose flux.
    • Grant Information:
      DK-02001 United States DK NIDDK NIH HHS; T32-HL-07260 United States HL NHLBI NIH HHS
    • Accession Number:
      0 (HSP70 Heat-Shock Proteins)
      0 (Hypoglycemic Agents)
      0 (Insulin)
      0 (Membrane Proteins)
      0 (Sp1 Transcription Factor)
      0 (Tubulin)
      IY9XDZ35W2 (Glucose)
      V956696549 (Acetylglucosamine)
    • Publication Date:
      Date Created: 20021025 Date Completed: 20030213 Latest Revision: 20131121
    • Publication Date:
      20231215
    • Accession Number:
      10.1152/ajpendo.00382.2002
    • Accession Number:
      12397027