Purification and preparation of Rhodobacter sphaeroides reaction centers for photocurrent measurements and atomic force microscopy characterization

Daniel Jun; Sylvester Zhang; Adrian Jan Grzędowski; Amita Mahey; J Thomas Beatty; Dan Bizzotto

doi:10.1016/j.xpro.2021.101044

Purification and preparation of Rhodobacter sphaeroides reaction centers for photocurrent measurements and atomic force microscopy characterization

STAR Protoc. 2021 Dec 16;3(1):101044. doi: 10.1016/j.xpro.2021.101044. eCollection 2022 Mar 18.

Authors

Daniel Jun^{1

2}, Sylvester Zhang^{1

3}, Adrian Jan Grzędowski^{1

3}, Amita Mahey², J Thomas Beatty², Dan Bizzotto^{1

3}

Affiliations

¹ Advanced Materials and Process Engineering Laboratory, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
² Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z1, Canada.
³ Department of Chemistry, University of British Columbia, Vancouver, BC V6T 1Z1, Canada.

Abstract

The formation of defined surfaces consisting of photosynthetic reaction centers (RCs) in biohybrid solar cells is challenging. Here, we start with the production of engineered RCs for oriented binding. RCs are deposited onto gold electrodes, and 6-mercapto-1-hexanol (MCH) is used to displace multilayers and non-specifically adsorbed RCs. The resulting electrode surfaces are analyzed for photocurrent generation using an intensity-modulated light and lock-in amplifier. Atomic force microscopy (AFM) is used to characterize the surface and the formation of RC structural assemblies. For complete details on the use and execution of this profile, please refer to Jun et al. (2021).

Keywords: Atomic Force Microscopy (AFM); Biophysics; Chemistry; Energy; Material sciences; Microbiology; Microscopy; Physics; Protein Biochemistry; Protein expression and purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Electrodes
Gold / chemistry
Microscopy, Atomic Force / methods
Photosynthetic Reaction Center Complex Proteins* / chemistry
Rhodobacter sphaeroides* / chemistry

Substances

Photosynthetic Reaction Center Complex Proteins
Gold