Synthetic Multidomain Proteins Containing Unstructured and α‑Helical Linkers Reveal a Differential Impact of Molecular Crowding on Catalytic Activity and Conformation.

Analyzing the BRET of both the 17 and 12 AAs-long unstructured linker-containing multidomain protein showed that smaller-molecular-weight PEGs (400, 2000, 4000, and 8000) showed a decreasing trend in BRET.

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The decrease in BRET of both the 17 and 12 AAs-long unstructured linker-containing multidomain protein in the presence of PEG 400 in comparison with the 21 AAs-long unstructured linker-containing multidomain protein showed a significant difference across all concentrations, with a faster decrease in BRET, while the 12 AAs-long multidomain protein in the presence of PEG 2000 showed an increasing trend ( Table S4 ).

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