Tandemization Endows Bovine Pancreatic Ribonuclease with Cytotoxic Activity

Leich, Franziska; Köditz, Jens; Ulbrich-Hofman, Renate; Arnold, Ulrich

doi:10.1016/j.jmb.2006.03.007

Cited by 27 publications

(41 citation statements)

References 47 publications

(73 reference statements)

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“…Moreover, the peptide linker allows to adjust the distance between the two RNase entities and to vary the charge of the molecule. Even though tandemization obviously did not result in an improved RI evasion in vitro, the constructs showed pronounced cytotoxicity (Leich et al 2006). An improved endocytosis as proposed for domain swapped RNase A oligomers (Libonati 2004) might be the reason for the increased cytotoxicity.…”

Section: Tandem Rnasesmentioning

confidence: 99%

“…Inspired by the dimeric structure of BS-RNase which impedes RI binding, Leich et al (2006) developed RNase A tandem enzymes where two RNase A molecules are fused head-to-tail by a peptide linker (Fig. 3).…”

Section: Tandem Rnasesmentioning

confidence: 99%

“…For many RNases or RNase variants cytotoxicity is proportional to RI evasion but other variants elude this relation. For instance, G88R/S90R-RNase 1 (Bosch et al 2004), domain-swapped RNase A dimers (Naddeo et al 2005), and RNase A tandem enzymes (Leich et al 2006) are bound and inactivated by RI in vitro but nevertheless they are cytotoxic. Inversely, the 8-fold RNase 1 mutant PI5 evades RI binding yet it is not cytotoxic (Bosch et al 2004).…”

Section: Open Questions and Concluding Remarksmentioning

confidence: 99%

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Natural and engineered ribonucleases as potential cancer therapeutics

Arnold

Ulbrich‐Hofmann

2006

Biotechnol Lett

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By reason of their cytotoxicity, ribonucleases (RNases) are potential anti-tumor drugs. Particularly members from the RNase A and RNase T1 superfamilies have shown promising results. Among these enzymes, Onconase, an RNase from the Northern Leopard frog, is furthest along in clinical trials. A general model for the mechanism of the cytotoxic action of RNases includes the interaction of the enzyme with the cellular membrane, internalization, translocation to the cytosol, and degradation of ribonucleic acid. The interplay of these processes as well as the role of the thermodynamic and proteolytic stability, the catalytic activity, and the capability of the RNase to evade the intracellular RNase inhibitor has not yet been fully elucidated. This paper discusses the various approaches to exploit RNases as cytotoxic agents.

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Section: Tandem Rnasesmentioning

confidence: 99%

Section: Tandem Rnasesmentioning

confidence: 99%

Section: Open Questions and Concluding Remarksmentioning

confidence: 99%

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Natural and engineered ribonucleases as potential cancer therapeutics

Arnold

Ulbrich‐Hofmann

2006

Biotechnol Lett

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“…To calculate values of [D] 50% , the concentration of denaturant at which 50% of the protein is unfolded, the signals y were fitted by nonlinear regression according to Santoro and Bolen [50] as described previously [38,51].…”

Section: Gdnhcl-induced Transition Curves and Determination Of The Pamentioning

confidence: 99%

The effect of additional disulfide bonds on the stability and folding of ribonuclease A

Pecher¹,

Arnold²

2009

Biophysical Chemistry

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International audienceThe significant contribution of disulfide bonds to the conformational stability of proteins is generally considered to result from an entropic destabilization of the unfolded state causing a faster escape of the molecules to the native state. However, the introduction of extra disulfide bonds into proteins as a general approach to protein stabilization yields rather inconsistent results. By modeling studies, we selected positions to introduce additional disulfide bonds into ribonuclease A at regions that had proven to be crucial for the initiation of the folding or unfolding process, respectively. However, only two out of the six variants proved to be more stable than unmodified ribonuclease A. The comparison of the thermodynamic and kinetic data disclosed a more pronounced effect on the unfolding reaction for all variants regardless of the position of the extra disulfide bond. Native-state proteolysis indicated a perturbation of the native state of the destabilized variants that obviously counterbalances the stability gain by the extra disulfide bond

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“…Results from control experiments in which RNase A was assayed in 100 mM Mes-NaOH (pH 6.0), 0.1 M NaCl were directly comparable to those obtained by other authors. [24][25][26] All other assays were Figure 1. Purification of Amphinases.…”

Section: Ribonucleolytic Activitymentioning

confidence: 99%