Raman microscopy is having increasing applications into molecular biology, ranging from studies on ligand binding [1] and secondary structure analysis [2]. Here we present results of a combined Raman microscopy and X-ray crystallography approach to study protein single crystals. The combined approach reinforces the interpretation of biophysical data. Both tetrameric hemoglobins and collagen-like polypeptides have been ivestigated. Tetrameric hemoglobins from Antarctic fish (AFHb) exibit a peculiar autoxidation pathway [3-5], that has been investigated via crystallography, and via Raman spectro/microscopy (both in solution and in crystal phase). The appearance of transient Raman bands both in solution and in crystals of AFHb at short time of air-exposure suggests that a bis-histidyl ferric adduct forms after the ferrous iron oxidizes to a penta-coordinated ferric state. A collagen-like polypeptide, (Pro-Pro-Gly)10, whose triple helix crystal structure has been solved at high resolution [6], has been also investigated. The comparison of amide I and amide III bands of the single crystal with the x-ray crystal structure of the triple helix provides an accurate reference to predict relationship between amide band frequencies and Ramachandran angles [7]. Acknowledgement We acknowledge Regione Campania and PNRA for financial support. References [1] P. R.Carey, J. Dong, Biochemistry 2004, 43, 8885-8893. [2] P. R. Carey, Ann. Rev. Phys. Chem. 2006, 57, 527-554. [3] A. Riccio, L. Vitagliano, G. di Prisco, A. Zagari, L. Mazzarella., Proc. Natl. Acad. Sci. USA 2002, 99, 9801-9806. [4] L. Vitagliano, G. Bonomi, A. Riccio, G. di Prisco, G. Smulevich, L. Mazzarella.. Eur. J. Biochem. 2004, 271, 1651-1659. [5] A. Vergara, M. Franzese, A. Merlino, L. Vitagliano, C. Verde, G. di Prisco, H. C. Lee, J. Peisach, L. Mazzarella, , Biophys. J. 2007, in press. [6] R. Berisio, L. Vitagliano, L. Mazzarella, A. Zagari,. Prot. Sci. 2002, 11, 262-270. [7] S.A. Asher, A. Ianoul, G. Mix, M. N. Boyden, A. Karnoup, M. Diem, R. Schweitzer-Stenner, J. Am. Chem. Soc. 2001, 123, 11775-11781.
A combined Raman microscopy and X-ray crystallography approach to study protein single crystals / Vergara, Alessandro; Merlino, Antonello; L., Vitagliano; Zagari, Adriana; Mazzarella, Lelio. - STAMPA. - (2007), pp. 152-152. (Intervento presentato al convegno XIIth European Conference on the Spectroscopy of Biological Molecules tenutosi a Bobigny, (FRANCIA) nel 1-6 settembre 2007).
A combined Raman microscopy and X-ray crystallography approach to study protein single crystals
VERGARA, ALESSANDRO;MERLINO, ANTONELLO;ZAGARI, ADRIANA;MAZZARELLA, LELIO
2007
Abstract
Raman microscopy is having increasing applications into molecular biology, ranging from studies on ligand binding [1] and secondary structure analysis [2]. Here we present results of a combined Raman microscopy and X-ray crystallography approach to study protein single crystals. The combined approach reinforces the interpretation of biophysical data. Both tetrameric hemoglobins and collagen-like polypeptides have been ivestigated. Tetrameric hemoglobins from Antarctic fish (AFHb) exibit a peculiar autoxidation pathway [3-5], that has been investigated via crystallography, and via Raman spectro/microscopy (both in solution and in crystal phase). The appearance of transient Raman bands both in solution and in crystals of AFHb at short time of air-exposure suggests that a bis-histidyl ferric adduct forms after the ferrous iron oxidizes to a penta-coordinated ferric state. A collagen-like polypeptide, (Pro-Pro-Gly)10, whose triple helix crystal structure has been solved at high resolution [6], has been also investigated. The comparison of amide I and amide III bands of the single crystal with the x-ray crystal structure of the triple helix provides an accurate reference to predict relationship between amide band frequencies and Ramachandran angles [7]. Acknowledgement We acknowledge Regione Campania and PNRA for financial support. References [1] P. R.Carey, J. Dong, Biochemistry 2004, 43, 8885-8893. [2] P. R. Carey, Ann. Rev. Phys. Chem. 2006, 57, 527-554. [3] A. Riccio, L. Vitagliano, G. di Prisco, A. Zagari, L. Mazzarella., Proc. Natl. Acad. Sci. USA 2002, 99, 9801-9806. [4] L. Vitagliano, G. Bonomi, A. Riccio, G. di Prisco, G. Smulevich, L. Mazzarella.. Eur. J. Biochem. 2004, 271, 1651-1659. [5] A. Vergara, M. Franzese, A. Merlino, L. Vitagliano, C. Verde, G. di Prisco, H. C. Lee, J. Peisach, L. Mazzarella, , Biophys. J. 2007, in press. [6] R. Berisio, L. Vitagliano, L. Mazzarella, A. Zagari,. Prot. Sci. 2002, 11, 262-270. [7] S.A. Asher, A. Ianoul, G. Mix, M. N. Boyden, A. Karnoup, M. Diem, R. Schweitzer-Stenner, J. Am. Chem. Soc. 2001, 123, 11775-11781.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
