Next Article in Journal
Structural Features of Heparan Sulfate from Multiple Osteochondromas and Chondrosarcomas
Previous Article in Journal
Four New Depsides Isolated from Salvia miltiorrhiza and Their Significant Nerve-Protective Activities
Article Menu
Issue 12 (December) cover image

Export Article

Open AccessArticle
Molecules 2018, 23(12), 3273; https://doi.org/10.3390/molecules23123273

Construction of Highly Stable Cytotoxic Nuclear-Directed Ribonucleases

1
Laboratori d’Enginyeria de Proteïnes, Departament de Biologia, Facultat de Ciències, Universitat de Girona, Campus de Montilivi, Maria Aurèlia Capmany 40, 17003 Girona, Spain
2
Institut d’Investigació Biomèdica de Girona Josep Trueta, (IdIBGi), 17190 Girona, Spain
*
Author to whom correspondence should be addressed.
Received: 8 November 2018 / Revised: 5 December 2018 / Accepted: 8 December 2018 / Published: 11 December 2018
(This article belongs to the Section Chemical Biology)
Full-Text   |   PDF [1543 KB, uploaded 11 December 2018]   |  

Abstract

Ribonucleases are proteins whose use is promising in anticancer therapy. We have previously constructed different human pancreatic ribonuclease variants that are selectively cytotoxic for tumor cells by introducing a nuclear localization signal into their sequence. However, these modifications produced an important decrease in their stability compromising their behavior in vivo. Here, we show that we can significantly increase the thermal stability of these cytotoxic proteins by introducing additional disulfide bonds by site-directed mutagenesis. One of these variants increases its thermal stability by around 17 °C, without affecting its catalytic activity while maintaining the cytotoxic activity against tumor cells. We also show that the most stable variant is significantly more resistant to proteolysis when incubated with proteinase K or with human sera, suggesting that its half-live could be increased in vivo once administered. View Full-Text
Keywords: antitumor protein; nuclear-directed ribonuclease; disulfide bond; thermal stability; proteolytic resistance antitumor protein; nuclear-directed ribonuclease; disulfide bond; thermal stability; proteolytic resistance
Figures

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Roura Padrosa, D.; Castro, J.; Romero-Casañas, A.; Ribó, M.; Vilanova, M.; Benito, A. Construction of Highly Stable Cytotoxic Nuclear-Directed Ribonucleases. Molecules 2018, 23, 3273.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top