Two types of amorphous protein particles facilitate crystal nucleation

Yamazaki Tomoya; Kimura Yuki; Vekilov Peter G.; Furukawa Erika; Shirai Manabu; Matsumoto Hiroaki; Van Driessche Alexander E. S.; Tsukamoto Katsuo

Title	en Two types of amorphous protein particles facilitate crystal nucleation
Creator	en Yamazaki, Tomoya en Kimura, Yuki NRID 1000050449542 en Vekilov, Peter G. en Furukawa, Erika en Shirai, Manabu en Matsumoto, Hiroaki en Van Driessche, Alexander E. S. en Tsukamoto, Katsuo
Accessrights	open access
Subject	Other en nucleation Other en protein Other en lysozyme Other en transmission electron microscopy Other en in situ observation
Description	Abstract en Nucleation, the primary step in crystallization, dictates the number of crystals, the distribution of their sizes, the polymorph selection, and other crucial properties of the crystal population. We used timeresolved liquid-cell transmission electron microscopy (TEM) to perform an in situ examination of the nucleation of lysozyme crystals. Our TEM images revealed that mesoscopic clusters, which are similar to those previously assumed to consist of a dense liquid and serve as nucleation precursors, are actually amorphous solid particles (ASPs) and act only as heterogeneous nucleation sites. Crystalline phases never form inside them. We demonstrate that a crystal appears within a noncrystalline particle assembling lysozyme on an ASP or a container wall, highlighting the role of heterogeneous nucleation. These findings represent a significant departure from the existing formulation of the two-step nucleation mechanism while reaffirming the role of noncrystalline particles. The insights gained may have significant implications in areas that rely on the production of protein crystals, such as structural biology, pharmacy, and biophysics, and for the fundamental understanding of crystallization mechanisms.
Publisher	en National Academy of Sciences.
Date	Issued2017-01-10
Language	eng
Resource Type	journal article
Version Type	AM
Identifier	HDL http://hdl.handle.net/2115/66517
Relation	isVersionOf DOI https://doi.org/10.1073/pnas.1606948114
Journal	PISSN 0027-8424 en Proceedings of the National Academy of Sciences of the United States of America (PNAS) Volume Number114 Issue Number9 Page Start2154 Page End2159
File	fulltext Main_PNAS_accepted -.pdf 1.7 MB (application/pdf) Issued2017-01-10
Oaidate	2023-07-26