An upper limit for macromolecular crowding effects

作者：Miklos, Andrew C.; Li, Conggang; Sorrell, Courtney D.; Lyon, L. Andrew; Pielak, Gary J.^*

来源：BMC Biophysics, 2011, 4: 13.

DOI：10.1186/2046-1682-4-13

• MICROGEL THIN-FILMS
• HYDROGEN-EXCHANGE
• PROTEIN STABILITY
• CHYMOTRYPSIN INHIBITOR
• BACKBONE DYNAMICS
• NMR RELAXATION
• MECHANISM
• RATES

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 similar to 2 nm when crowded by a synthetic microgel composed of poly(N-isopropylacrylamide-co-acrylic acid) with particle radii of similar to 300 nm.
Results: Our studies revealed no change in protein rotational or ps-ns backbone dynamics and only mild (similar to 0.5 kcal/mol at 37 degrees 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.

• 出版日期2011-5-31
• 单位中国科学院武汉物理与数学研究所