Hostname: page-component-78c5997874-8bhkd Total loading time: 0 Render date: 2024-11-03T02:45:08.286Z Has data issue: false hasContentIssue false

Understanding Nanoscale Interfaces to the Proteins Cytochrome c and Ribonuclease S

Published online by Cambridge University Press:  12 July 2019

Kimberly Hamad-Schifferli*
Affiliation:
Mechanical Engineering, Biological Engineering, MIT
Get access

Abstract

Format

This is a copy of the slides presented at the meeting but not formally written up for the volume.

Abstract

Nanoscale interfaces to proteins have been achieved for a variety of applications, in the form of electrodes to measure conductivity, for sensing on cantilevers or fluorescent quantum dots, or nanoparticles that can be used as reporters in receptor-ligand binding assays. One prevailing requirement is that the biological function of the protein is maintained when linked to nanoscale systems. Due to the structure-function relationship of proteins, the protein must maintain its folded structure. We covalently link cytochrome c and Ribonuclease S to Au or magnetic nanoparticles (NPs) and study the interface, with the goal of constructing design rules that govern the interaction. In both cases we devise methods to achieve linkage of a nanoparticle to the protein on a specific amino acid, in addition to chemical treatments that minimize non-specific adsorption. The protein linked to the nanoparticles is biophysically characterized. Protein secondary structure was quantified by circular dichroism spectroscopy (CD). From these measurements we determine that electrostatic forces dominate the NP-protein interaction and minimization of these results in folded proteins with minimal non-specific adsorption. For Ribonuclease S, these findings are integrated with measurements of enzymatic activity and binding constants KM to yield a picture of the how the protein interaction with the NP affects its binding to the substrate and activity. Experiments in which the NP labeling position, NP ligand, size, and material (Au, Fe3O4, CoFe2O4) are systematically varied will be discussed.

Type
Slide Presentations
Copyright
Copyright © Materials Research Society 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)