title: Optimized biorecognition of cytochrome c 551 and azurin immobilized on thiol-terminated monolayers assembled on Au(111) substrates.
abstract: Molecular recognition between two redox partners, azurin and cytochrome c 551, is studied at the single-molecule level by means of atomic force spectroscopy, after optimizing azurin adsorption on gold via sulfhydryl-terminated alkanethiol spacers. Our experiments provide evidence of specific interaction between the two partners, thereby demonstrating that azurin preserves biorecognition capability when assembled on gold via these spacers. Additionally, the measured single-molecule kinetic reaction rate results are consistent with a likely transient nature of the complex. Interestingly, the immobilization strategy adopted here, which was previously demonstrated to favor electrical coupling between azurin (AZ) and the metal electrode, is also found to facilitate AZ interaction with the redox partner, if compared to the case of AZ directly adsorbed on bare gold. Our findings confirm the key role of a well-designed immobilization strategy, capable of optimizing both biorecognition capabilities and electrical coupling with the conductive substrate at the single-molecule level, as a starting point for advanced applications of redox proteins for ultrasensitive biosensing.
authors.affiliation: Biophysics and Nanoscience Centre, CNISM, and CNR-INFM, Dipartimento di Scienze Ambientali, Università della Tuscia, Largo dell'Università, I-01100 Viterbo, Italy. bonanni@unitus.it
authors.forename: B
authors.initials: B
authors.lastname: Bonanni
authors.forename: A R
authors.initials: AR
authors.lastname: Bizzarri
authors.forename: S
authors.initials: S
authors.lastname: Cannistraro
mesh_terms: D001400:Azurin / Q000032:analysis / Q000737:chemistry; D015374:Biosensing Techniques; D003578:Cytochrome-c Peroxidase / Q000032:analysis / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D010084:Oxidation-Reduction; D011550:Pseudomonas aeruginosa / Q000201:enzymology; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D013438:Sulfhydryl Compounds; D001400:Azurin; D006046:Gold; D003578:Cytochrome-c Peroxidase