TY - JOUR

T1 - Molecular-level insights into the surface-induced assembly of Functional Bacterial Amyloid

AU - Sønderby, Thorbjørn Vincent

AU - Zou, Yimin

AU - Wang, Pengyu

AU - Wang, Chen

AU - Otzen, Daniel Erik

N1 - Copyright © 2022 Biophysical Society. Published by Elsevier Inc. All rights reserved.

PY - 2022/9

Y1 - 2022/9

N2 - Protein coating material is important in many technological fields. The interaction between carbon nanomaterial and protein is especially interesting since it makes the development of novel hybrid materials possible. Functional Bacterial Amyloid (FuBA) is promising as a coating material because of its desirable features such as well-defined molecular structure, robustness against harsh conditions, and easily engineerable functionality. Here, we report the systematic assembly of the functional amyloid protein, CsgA from Escherichia coli (E. coli) on graphite. We characterize the assemblies using Scanning Tunnelling Microscopy (STM) and show that CsgA forms assemblies according to systematic patterns, dictated by the graphite lattice. In addition, we show that graphite flakes induce the fibrillization of CsgA, in vitro, suggesting a surface-induced conformational change of CsgA facilitated by the graphite lattice. Using coarse-grained molecular dynamics simulations, we model the adhesion and lamellar formation of a CsgA-derived peptide and conclude that peptides are adsorbed both as monomers and smaller aggregates leading initially to unordered graphite-bound aggregates which are followed by rearrangement into lamellar structures. Finally, we show that CsgA-derived peptides can be immobilized in very systematic assemblies and their molecular orientation can be tuned using a small chaperone-like molecule. Our findings have implications for the development of FuBA-based biosensors, catalysts, and other technologies requiring well-defined protein assemblies on graphite.

AB - Protein coating material is important in many technological fields. The interaction between carbon nanomaterial and protein is especially interesting since it makes the development of novel hybrid materials possible. Functional Bacterial Amyloid (FuBA) is promising as a coating material because of its desirable features such as well-defined molecular structure, robustness against harsh conditions, and easily engineerable functionality. Here, we report the systematic assembly of the functional amyloid protein, CsgA from Escherichia coli (E. coli) on graphite. We characterize the assemblies using Scanning Tunnelling Microscopy (STM) and show that CsgA forms assemblies according to systematic patterns, dictated by the graphite lattice. In addition, we show that graphite flakes induce the fibrillization of CsgA, in vitro, suggesting a surface-induced conformational change of CsgA facilitated by the graphite lattice. Using coarse-grained molecular dynamics simulations, we model the adhesion and lamellar formation of a CsgA-derived peptide and conclude that peptides are adsorbed both as monomers and smaller aggregates leading initially to unordered graphite-bound aggregates which are followed by rearrangement into lamellar structures. Finally, we show that CsgA-derived peptides can be immobilized in very systematic assemblies and their molecular orientation can be tuned using a small chaperone-like molecule. Our findings have implications for the development of FuBA-based biosensors, catalysts, and other technologies requiring well-defined protein assemblies on graphite.

KW - Amyloid/chemistry

KW - Amyloidogenic Proteins/metabolism

KW - Escherichia coli Proteins/metabolism

KW - Escherichia coli/metabolism

KW - Graphite

KW - Peptides/chemistry

U2 - 10.1016/j.bpj.2022.08.013

DO - 10.1016/j.bpj.2022.08.013

M3 - Journal article

C2 - 35982614

VL - 121

SP - 3422

EP - 3434

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 18

ER -