title: Effects of spermine NONOate and ATP on the thermal stability of hemoglobin.
abstract: BACKGROUND
  Minor changes in protein structure induced by small organic and inorganic molecules can result in significant metabolic effects. The effects can be even more profound if the molecular players are chemically active and present in the cell in considerable amounts. The aim of our study was to investigate effects of a nitric oxide donor (spermine NONOate), ATP and sodium/potassium environment on the dynamics of thermal unfolding of human hemoglobin (Hb). The effect of these molecules was examined by means of circular dichroism spectrometry (CD) in the temperature range between 25°C and 70°C. The alpha-helical content of buffered hemoglobin samples (0.1 mg/ml) was estimated via ellipticity change measurements at a heating rate of 1°C/min.
  
  
  RESULTS
  Major results were: 1) spermine NONOate persistently decreased the hemoglobin unfolding temperature Tuirrespectively of the Na + /K + environment, 2) ATP instead increased the unfolding temperature by 3°C in both sodium-based and potassium-based buffers and 3) mutual effects of ATP and NO were strongly influenced by particular buffer ionic compositions. Moreover, the presence of potassium facilitated a partial unfolding of alpha-helical structures even at room temperature.
  
  
  CONCLUSIONS
  The obtained data might shed more light on molecular mechanisms and biophysics involved in the regulation of protein activity by small solutes in the cell.
authors.affiliation: Institute of Bioengineering (IFB), Aachen University of Applied Sciences, 52428 Juelich, Germany. bassam@fh-aachen.de.
authors.forename: Rasha
authors.initials: R
authors.lastname: Bassam
authors.forename: Juergen
authors.initials: J
authors.lastname: Hescheler
authors.forename: Ayseguel
authors.initials: A
authors.lastname: Temiz-Artmann
authors.forename: Gerhard M
authors.initials: GM
authors.lastname: Artmann
authors.forename: Ilya
authors.initials: I
authors.lastname: Digel