title: Metal-activated C-peptide facilitates glucose clearance and the release of a nitric oxide stimulus via the GLUT1 transporter.
abstract: OBJECTIVE
  Proinsulin C-peptide has been implicated in reducing complications associated with diabetes and also in improving blood flow. We hypothesised that incubation of erythrocytes with C-peptide would improve the ability of these cells to release ATP, a stimulus for nitric oxide production.
  
  
  METHODS
  Erythrocytes obtained from rabbits (n = 11) and both healthy and type 2 diabetic humans (n = 7) were incubated with C-peptide in the absence and presence of Fe2+ and Cr3+, and the resulting ATP release was measured via chemiluminescence. This release was also measured in the presence and absence of phloretin, an inhibitor of GLUT1, and also of mannose, a glycolysis inhibitor. To determine glucose transport, 14C-labelled glucose was added to erythrocytes in the presence and absence of the C-peptide-metal complex and the aforementioned inhibitors.
  
  
  RESULTS
  The release of ATP from the erythrocytes of patients with diabetes increased from 64 +/- 13 to 260 +/- 39 nmol/l upon incubation of the cells in C-peptide. The C-peptide activity was dependent upon binding to Fe2+, which was extended upon binding to Cr3+. The increase in ATP release from the erythrocytes is due to metal-activated C-peptide stimulation of glucose transfer into the erythrocytes via the GLUT1 transporter. In the presence of C-peptide complexed to Cr3+, the amount of glucose transferred into the erythrocyte increased by 31%.
  
  
  CONCLUSIONS
  When complexed to Fe2+ or Cr3+, C-peptide has the ability to promote ATP release from erythrocytes. This release is due to an increase in glucose transport through GLUT1.
authors.affiliation: Department of Chemistry, Wayne State University, 5101 Cass Avenue, Detroit, MI 48202, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Meyer
authors.forename: J M
authors.initials: JM
authors.lastname: Froelich
authors.forename: G E
authors.initials: GE
authors.lastname: Reid
authors.forename: W K A
authors.initials: WK
authors.lastname: Karunarathne
authors.forename: D M
authors.initials: DM
authors.lastname: Spence
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002096:C-Peptide / Q000378:metabolism*; D002857:Chromium / Q000737:chemistry; D004912:Erythrocytes / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D051272:Glucose Transporter Type 1 / Q000378:metabolism*; D006019:Glycolysis; D006801:Humans; D007501:Iron / Q000737:chemistry; D008297:Male; D008670:Metals / Q000378:metabolism*; D008954:Models, Biological; D009569:Nitric Oxide / Q000378:metabolism*; D011817:Rabbits
chemical_list: D002096:C-Peptide; D051272:Glucose Transporter Type 1; D008670:Metals; D002857:Chromium; D009569:Nitric Oxide; D000255:Adenosine Triphosphate; D007501:Iron; D005947:Glucose