European Biophysics Journal

, Volume 39, Issue 8, pp 1129–1142 | Cite as

Spectral characterization of the recombinant mouse tumor suppressor 101F6 protein

  • Alajos Bérczi
  • Filip Desmet
  • Sabine Van Doorslaer
  • Han Asard
Original Paper


Tumor suppressor protein 101F6, a gene product of the 3p21.3 (human) and 9F1 (mouse) chromosomal region, has recently been identified as a member of the cytochrome b561 (Cyt-b561) protein family by sequence homology. The His6-tagged recombinant mouse tumor suppressor Cyt-b561 protein (TSCytb) was recently expressed in yeast and purified, and the ascorbate reducibility was determined. TSCytb is auto-oxidizable and has two distinct heme b centers with redox potentials of ~40 and ~140 mV. Its split α-band in the dithionite-reduced spectrum at both 295 and 77 K is well resolved, and the separation between the two α-peaks is ~7 nm (~222 cm−1). Singular value decomposition analysis of the split α-band in the ascorbate-reduced spectra revealed the presence of two major spectral components, each of them with split α-band but with different peak separations (6 and 8 nm). Similar minor differences in peak separation were obtained when the split α-bands in ascorbate-reduced difference spectra at low (<1 mM) and high (>10 mM) ascorbate concentrations were analysed. According to low-temperature electron paramagnetic resonance (EPR) spectroscopy, the two heme b centers are in the low-spin ferric state with maximum principal g values of 3.61 and 2.96, respectively. These values differ from the ones observed for other members of the Cyt-b561 family. According to resonance Raman spectroscopy, the porphyrin rings are in a relaxed state. The spectroscopic results are only partially in agreement with those obtained earlier for the native chromaffin granule Cyt-b561.


Ascorbate Auto-oxidation Cyt-b561 protein EPR Raman UV–VIS 101F6 protein 



The authors thank Drs. Balázs Szalontai and Csaba Bagyinka (Institute of Biophysics, BRC, Szeged, Hungary) for their help in spectrum analysis. This work was supported by grants from the University of Antwerp (to H.A.). F.D. thanks the BOF-UA-TOP fund for PhD funding.


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Copyright information

© European Biophysical Societies' Association 2009

Authors and Affiliations

  • Alajos Bérczi
    • 1
  • Filip Desmet
    • 2
  • Sabine Van Doorslaer
    • 2
  • Han Asard
    • 3
  1. 1.Institute of Biophysics, Biological Research CenterHungarian Academy of SciencesSzegedHungary
  2. 2.Department of PhysicsUniversity of AntwerpWilrijkBelgium
  3. 3.Department of BiologyUniversity of AntwerpAntwerpenBelgium

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