Email updates

Keep up to date with the latest news and content from BMC Biophysics and BioMed Central.

Open Access Highly Accessed Research article

An upper limit for macromolecular crowding effects

Andrew C Miklos1, Conggang Li12, Courtney D Sorrell345, L Andrew Lyon34 and Gary J Pielak167*

Author Affiliations

1 Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina 27599, USA

2 State Key Laboratory of Magnetic Resonance and Molecular and Atomic Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, 430071, PR China

3 School of Chemistry & Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA

4 Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA

5 Department of Chemistry, University of Alberta, Edmonton, AB, T6G 2G2, Canada

6 Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina 27599, USA

7 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA

For all author emails, please log on.

BMC Biophysics 2011, 4:13  doi:10.1186/2046-1682-4-13

Published: 31 May 2011

Abstract

Background

Solutions containing high macromolecule concentrations are predicted to affect a number of protein properties compared to those properties in dilute solution. In cells, these macromolecular crowders have a large range of sizes and can occupy 30% or more of the available volume. We chose to study the stability and ps-ns internal dynamics of a globular protein whose radius is ~2 nm when crowded by a synthetic microgel composed of poly(N-isopropylacrylamide-co-acrylic acid) with particle radii of ~300 nm.

Results

Our studies revealed no change in protein rotational or ps-ns backbone dynamics and only mild (~0.5 kcal/mol at 37°C, pH 5.4) stabilization at a volume occupancy of 70%, which approaches the occupancy of closely packing spheres. The lack of change in rotational dynamics indicates the absence of strong crowder-protein interactions.

Conclusions

Our observations are explained by the large size discrepancy between the protein and crowders and by the internal structure of the microgels, which provide interstitial spaces and internal pores where the protein can exist in a dilute solution-like environment. In summary, microgels that interact weakly with proteins do not strongly influence protein dynamics or stability because these large microgels constitute an upper size limit on crowding effects.