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Spectral characterization of the recombinant mouse tumor suppressor 101F6 protein.

Abstract
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 His(6)-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 approximately 40 and approximately 140 mV. Its split alpha-band in the dithionite-reduced spectrum at both 295 and 77 K is well resolved, and the separation between the two alpha-peaks is approximately 7 nm (approximately 222 cm(-1)). Singular value decomposition analysis of the split alpha-band in the ascorbate-reduced spectra revealed the presence of two major spectral components, each of them with split alpha-band but with different peak separations (6 and 8 nm). Similar minor differences in peak separation were obtained when the split alpha-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.
AuthorsAlajos Bérczi, Filip Desmet, Sabine Van Doorslaer, Han Asard
JournalEuropean biophysics journal : EBJ (Eur Biophys J) Vol. 39 Issue 8 Pg. 1129-42 (Jul 2010) ISSN: 1432-1017 [Electronic] Germany
PMID19943161 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Cyb561d2 protein, mouse
  • Cytochrome b Group
  • Membrane Proteins
  • Micelles
  • Recombinant Proteins
  • Tumor Suppressor Proteins
  • Dithionite
  • Heme
Topics
  • Amino Acid Sequence
  • Animals
  • Cytochrome b Group (chemistry, genetics)
  • Dithionite
  • Electron Spin Resonance Spectroscopy (methods)
  • Heme (chemistry)
  • Membrane Proteins
  • Mice
  • Micelles
  • Oxidation-Reduction
  • Recombinant Proteins (chemistry, genetics)
  • Saccharomyces
  • Sequence Homology, Amino Acid
  • Spectrum Analysis (methods)
  • Spectrum Analysis, Raman
  • Temperature
  • Tumor Suppressor Proteins (chemistry, genetics)
  • Ultraviolet Rays

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