TY - JOUR
T1 - A new method to determine the structure of the metal environment in metalloproteins
T2 - Investigation of the prion protein octapeptide repeat Cu 2+ complex
AU - Mentler, Matthias
AU - Weiss, Andreas
AU - Grantner, Klaus
AU - Del Pino, Pablo
AU - Deluca, Dominga
AU - Fiori, Stella
AU - Renner, Christian
AU - Klaucke, Wolfram Meyer
AU - Moroder, Luis
AU - Bertsch, Uwe
AU - Kretzschmar, Hans A.
AU - Tavan, Paul
AU - Parak, Fritz G.
N1 - Funding Information:
Acknowledgements This work was supported by the Bayerischer Forschungsverbund Prionen and the Bundesministerium für Bil-dung und Forschung.
PY - 2005/3
Y1 - 2005/3
N2 - Since high-intensity synchrotron radiation is available, "extended X-ray absorption fine structure" spectroscopy (EXAFS) is used for detailed structural analysis of metal ion environments in proteins. However, the information acquired is often insufficient to obtain an unambiguous picture. ENDOR spectroscopy allows the determination of hydrogen positions around a metal ion. However, again the structural information is limited. In the present study, a method is proposed which combines computations with spectroscopic data from EXAFS, EPR, electron nuclear double resonance (ENDOR) and electron spin echo envelope modulation (ESEEM). From EXAFS a first picture of the nearest coordination shell is derived which has to be compatible with EPR data. Computations are used to select sterically possible structures, from which in turn structures with correct H and N positions are selected by ENDOR and ESEEM measurements. Finally, EXAFS spectra are re-calculated and compared with the experimental data. This procedure was successfully applied for structure determination of the Cu2+ complex of the octapeptide repeat of the human prion protein. The structure of this octarepeat complex is rather similar to a pentapeptide complex which was determined by X-ray structure analysis. However, the tryptophan residue has a different orientation: the axial water is on the other side of the Cu.
AB - Since high-intensity synchrotron radiation is available, "extended X-ray absorption fine structure" spectroscopy (EXAFS) is used for detailed structural analysis of metal ion environments in proteins. However, the information acquired is often insufficient to obtain an unambiguous picture. ENDOR spectroscopy allows the determination of hydrogen positions around a metal ion. However, again the structural information is limited. In the present study, a method is proposed which combines computations with spectroscopic data from EXAFS, EPR, electron nuclear double resonance (ENDOR) and electron spin echo envelope modulation (ESEEM). From EXAFS a first picture of the nearest coordination shell is derived which has to be compatible with EPR data. Computations are used to select sterically possible structures, from which in turn structures with correct H and N positions are selected by ENDOR and ESEEM measurements. Finally, EXAFS spectra are re-calculated and compared with the experimental data. This procedure was successfully applied for structure determination of the Cu2+ complex of the octapeptide repeat of the human prion protein. The structure of this octarepeat complex is rather similar to a pentapeptide complex which was determined by X-ray structure analysis. However, the tryptophan residue has a different orientation: the axial water is on the other side of the Cu.
KW - Electron nuclear double resonance
KW - Electron spin echo envelope modulation
KW - Extended X-ray absorption fine structure spectroscopy
KW - Prion protein
KW - Protein structure
UR - http://www.scopus.com/inward/record.url?scp=20144387902&partnerID=8YFLogxK
U2 - 10.1007/s00249-004-0434-z
DO - 10.1007/s00249-004-0434-z
M3 - Article
C2 - 15452673
AN - SCOPUS:20144387902
SN - 0175-7571
VL - 34
SP - 97
EP - 112
JO - European Biophysics Journal
JF - European Biophysics Journal
IS - 2
ER -