Crowded, cell-like environment induces shape changes in aspherical protein
2009 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 96, no 3, Supplement 1, 568a- p.Article in journal (Refereed) Published
Protein dynamics in cells may be different from that in dilute solutions in vitro since the environment in cells is highly concentrated with other macromolecules. This volume exclusion due to macromolecular crowding is predicted to affect both equilibrium and kinetic processes involving protein conformational changes. To quantify macromolecular crowding effects on protein folding mechanisms, here we have investigated the folding energy landscape of an α/β protein, apoflavodoxin, in the presence of inert macromolecular crowding agents using in silico and in vitro approaches. By coarse-grained molecular simulations and topology-based potential interactions, we probed the effects of increased volume fraction of crowding agents (fc) as well as of crowding agent geometry (sphere or spherocylinder) at high fc. Parallel kinetic folding experiments with purified Desulfovibro desulfuricans apoflavodoxin in vitro were performed in the presence of Ficoll (sphere) and Dextran (spherocylinder) synthetic crowding agents. In conclusion, we have identified in silico crowding conditions that best enhance protein stability and discovered that upon manipulation of the crowding conditions, folding routes experiencing topological frustrations can be either enhanced or relieved. The test-tube experiments confirmed that apoflavodoxin's time-resolved folding path is modulated by crowding agent geometry. We propose that macromolecular crowding effects may be a tool for manipulation of protein folding and function in living cells.
Place, publisher, year, edition, pages
New York: Rockefeller U.P. for the Biophysical society , 2009. Vol. 96, no 3, Supplement 1, 568a- p.
IdentifiersURN: urn:nbn:se:umu:diva-22203DOI: 10.1016/j.bpj.2008.12.3723OAI: oai:DiVA.org:umu-22203DiVA: diva2:213651
Presentation vid Biophysical Society meeting i Boston i februari. Platform BE: Protein Folding & Stability II2009-04-282009-04-282011-04-12Bibliographically approved