Crystallization and preliminary X-ray analysis of the complex of human alpha-thrombin with a modified thrombin-binding aptamer

Crystallization and preliminary X-ray analysis of the complex of human alpha-thrombin with a modified thrombin-binding aptamer (267 visite)

Russo Krauss I, Merlino A, Randazzo A, Mazzarella L, Sica F

Nucleosides Nucleotides Nucleic Acids (ISSN: 1744-3091, 1744-4309, 1532-2335electronic), 2010 Aug 1; 66(Pt8): 961-963.

Tipo di articolo: Journal Article, Abstract, Conference,

Impact factor: 0.563, Impact factor a 5 anni: 0.479

Url: http://www.scopus.com/inward/record.url?eid=2-s2.0-77955446114&partnerID=40&md5=a4c2092a293437c2d79737ffbfa18074

Parole chiave: Protein-Nucleic Acid Interactions, Quadruplexes, Thrombin, Thrombin-Binding Aptamers, Thrombin Aptamer, Article, Chemistry, Crystallization, Human, Metabolism, Protein Binding, Protein Multimerization, X Ray Crystallography, Nucleotide, X-Ray, Base Sequence, Biomimetic Materials, Chemical Synthesis, Chemistry Techniques, Synthetic, Catalytic, Genetics, 10-23, Chimeric Oligonucleotide, Dnazyme, Deoxyribozyme, Nucleic Acid Crystallization,

Affiliazioni:

*** IBB - CNR ***
Dipartimento di Chimica, Universit di Napoli Federico II, Via Cintia, I-80126 Napoli, Italy
Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, I-80134 Napoli, Italy
a Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences , Department of Bioorganic Chemistry , Lodz , Poland.,

Riferimenti:

Bock, L. C., Griffin, L. C., Latham, J. A., Vermaas, E. H., Toole, J. J., (1992) Nature (London), 355, pp. 564-56

Bode, W., Mayr, I., Baumann, U., Huber, R., Stone, S. R., Hofsteenge, J., (1989) EMBO J., 8, pp. 3467-3475

Kelly, J. A., Feigon, J., Yeates, T. O., (1996) J. Mol. Biol., 256, pp. 417-422

Lancellotti, S., De Cristofaro, R., (2009) Cardiovasc. Hematol. Agents Med. Chem., 7, pp. 19-28

MacAya, R. F., Schultze, P., Smith, F. W., Roe, J. A., Feigon, J., (1993) Proc. Natl Acad. Sci. USA, 90, pp. 3745-3749

Martino, L., Virno, A., Randazzo, A., Virgilio, A., Esposito, V., Giancola, C., Bucci, M., Mayol, L., (2006) Nucleic Acids Res., 34, pp. 6653-6662

Navaza, J., Saludjian, P., (1997) Methods Enzymol., 276, pp. 581-594

Otwinowski, Z., Minor, W., (1997) Methods Enzymol., 276, pp. 307-326

Padmanabhan, K., Padmanabhan, K. P., Ferrara, J. D., Sadler, J. E., Tulinsky, A., (1993) J. Biol. Chem., 268, pp. 17651-17654

Padmanabhan, K., Tulinsky, A., (1996) Acta Cryst., D52, pp. 272-282

Pagano, B., Martino, L., Randazzo, A., Giancola, C., (2007) Biophys. J., 94, pp. 562-569

Schultze, P., MacAya, R. F., Feigon, J., (1994) J. Mol. Biol., 235, pp. 1532-1547

The thrombin-binding aptamer (TBA) is a consensus DNA 15-mer that binds specifically to human-thrombin at nanomolar concentrations and inhibits its procoagulant functions. Recently, a modified TBA (mTBA) containing a 5 -5 inversion-of-polarity site has been shown to be more stable and to possess a higher thrombin affinity than its unmodified counterpart. The structure of the thrombin-TBA complex has previously been determined at low resolution, but did not provide a detailed picture of the aptamer conformation or of the protein-DNA assembly, while that of the complex with mTBA is unknown. Crystallographic analysis of the thrombin-mTBA complex has been attempted. The crystals diffracted to 2. 15 resolution and belonged to space group I222. 2010 International Union of Crystallography All rights reserved