Grabbing yeast iso-1-cytochrome c by Cys102: an effective approach for the assembly of functionally active metalloprotein carpets

Langmuir : the ACS Journal of Surfaces and Colloids
Mimmo GerundaPaolo Facci

Abstract

We report an approach for immobilizing iso-1-cytochrome c from Saccharomyces cerevisiae on oxygen exposing surfaces derivatized with SH-terminated silanes. The SH moieties from silanes have been brought to react with the partially buried Cys102, forming an intermolecular disulfide bond which anchored covalently cytochrome c to the surface. The presence of a single cysteine residue on the protein surface imparted a well-defined orientation to the molecular edifice. Molecular constructs obtained with native cytochrome c and with a cysteine-depleted mutant (C102T) have been investigated by means of scanning force microscopy under liquid, which was performed to assay the quality of the molecular carpet, showing that the native protein formed a robust monolayer at the surface, whereas only a negligible amount of physisorbed molecules were detected in the case of a mutant. UV-vis absorption spectroscopy was performed to confirm that immobilization takes place via the Cys102 residue. Linear sweep voltammetric measurements showed retention of the redox activity of the covalently immobilized cytochrome c, confirming the viability of the proposed immobilization method for obtaining monolayers of redox active molecules.

References

Jan 10, 2002·The Journal of Biological Chemistry·Esa K J TuominenPaavo K J Kinnunen
May 9, 2002·Journal of the American Chemical Society·Gianantonio BattistuzziMarco Sola
Jul 31, 2002·Archives of Biochemistry and Biophysics·Gianantonio BattistuzziMarco Sola
Mar 1, 1959·The Biochemical Journal·E MARGOLIASH, N FROHWIRT
Oct 31, 2003·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Gianantonio BattistuzziMarco Sola

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Citations

Sep 27, 2006·Physical Chemistry Chemical Physics : PCCP·Andrea AlessandriniPaolo Facci
Feb 6, 2007·Chemistry and Physics of Lipids·Andrea AlessandriniUmberto Muscatello
May 28, 2005·Yeast
Oct 11, 2016·Chemical Society Reviews·Stefano CasaliniFabio Biscarini
Jan 16, 2008·The Journal of Physical Chemistry. B·Stefano CasaliniMarco Sola
Feb 25, 2009·ACS Applied Materials & Interfaces·Marialuisa CaiazzoPaolo Facci
Apr 20, 2006·Journal of the American Chemical Society·Carlo Augusto BortolottiMarco Sola

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