Role of clusters in nonclassical nucleation and growth of protein crystals.
The development of multistep nucleation theory has spurred on experimentalists to find intermediate metastable states that are relevant to the solidification pathway of the molecule under interest. A great deal of studies focused on characterizing the so-called "precritical clusters" that may arise in the precipitation process. However, in macromolecular systems, the role that these clusters might play in the nucleation process and in the second stage of the precipitation process, i.e., growth, remains to a great extent unknown. Therefore, using biological macromolecules as a model system, we have studied the mesoscopic intermediate, the solid end state, and the relationship that exists between them. We present experimental evidence that these clusters are liquid-like and stable with respect to the parent liquid and metastable compared with the emerging crystalline phase. The presence of these clusters in the bulk liquid is associated with a nonclassical mechanism of crystal growth and can trigger a self-purifying cascade of impurity-poisoned crystal surfaces. These observations demonstrate that there exists a nontrivial connection between the growth of the macroscopic crystalline phase and the mesoscopic intermediate which should not be ignored. On the other hand, our experimental data also show that clusters existing in protein solutions can significantly increase the nucleation rate and therefore play a relevant role in the nucleation process.
Structural Biology Brussels, Flanders Interuniversity Institute for Biotechnology, and Vrije Universiteit Brussel, 1050 Elsene, Belgium.
Mike
M
Sleutel
Alexander E S
AE
Van Driessche
D019747:Aldose-Ketose Isomerases / Q000737:chemistry; D003460:Crystallization; D006863:Hydrogen-Ion Concentration; D018613:Microscopy, Confocal; D008956:Models, Chemical; D009113:Muramidase / Q000737:chemistry; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry*; D012996:Solutions
D011506:Proteins; D012996:Solutions; D009113:Muramidase; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase
phase transition; prenucleation clusters; self-purification