Single molecule recognition between Cytochrome C 551 and gold-immobilized Azurin by force spectroscopy

被引：74

作者：

Bonanni, B

Kamruzzahan, ASM

Bizzarri, AR

Rankl, C

Gruber, HJ

Hinterdorfer, P

Cannistraro, S

机构：

[1] Univ Tuscia, Dipartimento Sci Ambientali, INFM, CNSIM,Biophys & Nanosci Ctr, I-01100 Viterbo, Italy

[2] Johannes Kepler Univ, A-4040 Linz, Austria

来源：

BIOPHYSICAL JOURNAL | 2005年 / 89卷 / 04期

基金：

奥地利科学基金会;

关键词：

D O I：

10.1529/biophysj.105.064097

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

Recent developments in single molecule force spectroscopy have allowed investigating the interaction between two redox partners, Azurin and Cytochrome C 551. Azurin has been directly chemisorbed on a gold electrode whereas cytochrome c has been linked to the atomic force microscopy tip by means of a heterobifunctional flexible cross-linker. When recording force-distance cycles, molecular recognition events could be observed, displaying unbinding forces of similar to 95 pN for an applied loading rate of 10 nN/s. The specificity of molecular recognition was confirmed by the significant decrease of unbinding probability observed in control block experiments performed adding free azurin solution in the fluid cell. In addition, the complex dissociation kinetics has been here investigated by monitoring the unbinding forces as a function of the loading rate: the thermal off-rate was estimated to be similar to 14 s(-1), much higher than values commonly estimated for complexes more stable than electron transfer complexes. Results here discussed represent the first studies on molecular recognition between two redox partners by atomic force microscopy.

引用

页码：2783 / 2791

页数：9

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