Structural features for the mechanism of antitumor action of a dimeric human pancreatic ribonuclease variant(351 views) Merlino A, Avella G, Di Gaetano S, Arciello A, Piccoli R, Mazzarella L, Sica F
Protein Sci (ISSN: 0961-8368, 1469-896xelectronic, 0961-8368linking), 2009 Jan; 18(1): 50-57.
Dipartimento di Chimica, Università degli Studi di Napoli Federico II, Via Cintia, Napoli 80126, Italy
Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, Napoli 80134, Italy
Dipartimento di Biologia Strutturale e Funzionale, Università degli Studi di Napoli Federico II, Via Cintia, Napoli 80126, Italy
Istituto Nazionale di Biostrutture e Biosistemi (INBB), Italy
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Leland, P. A., Raines, R. T., Cancer chemotherapy-ribonucleases to the rescue (2001) Chem Biol, 8, pp. 405-413
Kover, K. E., Bruix, M., Santoro, J., Batta, G., Laurents, D. V., Rico, M., The solution structure and dynamics of human pancreatic ribonuclease determined by NMR Spectroscopy provide insight into its remarkable biological activities and inhibition (2008) J Mol Biol, 379, pp. 953-965
Johnson, R. J., McCoy, J. G., Bingman, C. A., Phillips Jr, G. N., Raines, R. T., Inhibition of human pancreatic ribonuclease by the human ribonuclease inhibitor protein (2007) J Mol Biol, 368, pp. 434-449
Lee, F. S., Shapiro, R., Vallee, B. L., Tight-binding inhibition of angiogenin and ribonuclease A by placental ribonuclease inhibitor (1989) Biochemistry, 28, pp. 225-230
French, G. S., Wilson, K. S., On the treatment of negative intensity observations (1978) Acta Crystallogr Sect A, 34, pp. 517-525
Brunger, A. T., Adams, P. D., Clore, G. M., DeLano, W. L., Gros, P., Grosse-Kunstleve, R. W., Jiang, J. S., Warren, G. L., Crystallography & NMR system: A new software suite for macromolecular structure determination (1998) Acta Crystallogr D Biol Crystallogr, 54 (PART 5), pp. 905-921
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Hooft, R. W., Vriend, G., Sander, C., Abola, E. E., Errors in protein structures (1996) Nature, 381, p. 272
Structural features for the mechanism of antitumor action of a dimeric human pancreatic ribonuclease variant
A specialized class of RNases shows a high cytotoxicity toward tumor cell lines, which is critically dependent on their ability to reach the cytosol and to evade the action of the ribonuclease inhibitor (RI). The cytotoxicity and antitumor activity of bovine seminal ribonuclease (BSRNase), which exists in the native state as an equilibrium mixture of a swapped and an unswapped dimer, are peculiar properties of the swapped form. A dimeric variant (HHP2-RNase) of human pancreatic RNase, in which the enzyme has been engineered to reproduce the sequence of BSRNase helix-II (Gln28 -> Leu, Arg31 -> Cys, Arg32 -> Cys, and Asn34 -> Lys) and to eliminate a negative charge on the surface (Glu111 -> Gly), is also extremely cytotoxic. Surprisingly, this activity is associated also to the unswapped form of the protein. The crystal structure reveals that on this molecule the hinge regions, which are highly disordered in the unswapped form of BSRNase, adopt a very well-defined conformation in both subunits. The results suggest that the two hinge peptides and the two Leu28 side chains may provide an anchorage to a transient noncovalent dimer, which maintains Cys31 and Cys32 of the two subunits in proximity, thus stabilizing a quaternary structure, similar to that found for the noncovalent swapped dimer of BSRNase, that allows the molecule to escape RI and/or to enhance the formation of the interchain disulfides.
Structural features for the mechanism of antitumor action of a dimeric human pancreatic ribonuclease variant