Real-time observation of nonclassical protein crystallization kinetics

Andrea Sauter, Felix Roosen-Runge, Fajun Zhang, Gudrun Lotze, Robert M.J. Jacobs, Frank Schreiber

Research output: Contribution to journalArticlepeer-review

74 Citations (SciVal)

Abstract

We present a real-time study of protein crystallization of bovine-lactoglobulin in the presence of CdCl2 using small-angle X-ray scattering and optical microscopy. From observing the crystallization kinetics, we propose the following multistep crystallization mechanism that is consistent with our data. In the first step, an intermediate phase is formed, followed by the nucleation of crystals within the intermediate phase. During this period, the number of crystals increases with time, but the crystal growth is slowed down by the surrounding dense intermediate phase due to the low mobility. In the next step, the intermediate phase is consumed by nucleation and slow growth, and the crystals are exposed to the dilute phase. In this stage, the number of crystals becomes nearly constant, whereas the crystals grow rapidly due to access to the free protein molecules in the dilute phase. This real-time study not only provides evidence for a two-step nucleation process for protein crystallization but also elucidates the role and the structural signature of the metastable intermediate phase in this process.

Original languageEnglish
Pages (from-to)1485-1491
Number of pages7
JournalJournal of the American Chemical Society
Volume137
Issue number4
DOIs
Publication statusPublished - 2015 Feb 4
Externally publishedYes

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