Capturing ultrafast energy flow of a heme protein in crowded milieu

Energy flow in biomolecules is a dynamic process vital for understanding health, disease, and applications in biotechnology and medicine. In crowded environments, where biomolecular functions are modulated, comprehending energy flow becomes crucial for accurately understanding cellular processes like signaling and subsequent functions. This study employs ultrafast transient absorption spectroscopy to demonstrate energy funneling from the photoexcited heme of bovine heart cytochrome c to the protein exterior, in the presence of common synthetic (Dextran 40, Ficoll 70, PEG 8 and Dextran 70) and protein-based (BSA and β-LG) crowders. The through-space energy transfer mode for ferric and the methionine rebinding mode for ferrous cytochrome c show the strongest solvent coupling. The heterogeneous behaviour of crowders, influenced by crowder-protein interactions and caging effects at certain higher concentrations, reveal diverse trends. Notably, protein crowders perturb all transport routes of vibrational energy transfer, causing delays in energy transfer processes. These findings provide significant insights into the basic tenets of energy flow, one of the most fundamental processes, in crowded cellular environments.