Journal of Physical Chemistry Letters
Journal of Physical Chemistry Letters
5 years ago

Direct Observation of Insulin Association Dynamics with Time-Resolved X-ray Scattering

Direct Observation of Insulin Association Dynamics with Time-Resolved X-ray Scattering
Denis Leshchev, Jiyun Hong, Irina Kosheleva, Dolev Rimmerman, Lin X. Chen, Darren J. Hsu
Biological functions frequently require protein–protein interactions that involve secondary and tertiary structural perturbation. Here we study protein–protein dissociation and reassociation dynamics in insulin, a model system for protein oligomerization. Insulin dimer dissociation into monomers was induced by a nanosecond temperature-jump (T-jump) of ∼8 °C in aqueous solution, and the resulting protein and solvent dynamics were tracked by time-resolved X-ray solution scattering (TRXSS) on time scales of 10 ns to 100 ms. The protein scattering signals revealed the formation of five distinguishable transient species during the association process that deviate from simple two-state kinetics. Our results show that the combination of T-jump pump coupled to TRXSS probe allows for direct tracking of structural dynamics in nonphotoactive proteins.

Publisher URL: http://dx.doi.org/10.1021/acs.jpclett.7b01720

DOI: 10.1021/acs.jpclett.7b01720

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