Structural Evolution of Metastable Protein Aggregates in the Presence of Trivalent Salt Studied by (V)SANS and SAXS.
We present a study of the structural evolution of protein aggregates formed in solutions of a globular protein, β-lactoglobulin (BLG), in the presence of YCl3. These aggregates are often observed before crystallization starts and they are metastable with respect to the crystalline phase. Here we focus on the characterization of the hierarchical structure of this intermediate phase and its temperature dependent structure evolution using a combination of (very) small angle neutron and X-ray scattering (VSANS, SANS, and SAXS). Results show that the hierarchical structure ranges from nanometer scale protein monomer, dimer and compact protein clusters to micrometer scale fractal protein aggregates. Upon cooling, the overall hierarchical structure is preserved, but the evolution of the internal structure within the aggregates is clearly visible: the monomer-monomer correlation peak reduces its intensity and disappears completely at lower temperatures, whereas the cluster-cluster correlation is enhanced. At a larger length scale, the fractal dimension of protein aggregates increases. The kinetics of the structure change during a temperature ramp was further investigated using time-resolved SAXS. The time dependent SAXS profiles show clear isosbestic points and the kinetics of the structural evolution can be well described using a two-state model. These dynamic properties of protein aggregates on a broad length scale may be essential for being the precursors of nucleation.
Institut für Angewandte Physik, Universität Tübingen , Auf der Morgenstelle 10, 72076 Tübingen, Germany.
Andrea
A
Sauter
Institut für Angewandte Physik, Universität Tübingen , Auf der Morgenstelle 10, 72076 Tübingen, Germany.
Fajun
F
Zhang
Forschungszentrum Jülich GmbH, JCNS @ MLZ , Lichtenbergstraße 1, 85747 Garching, Germany.
Noemi K
NK
Szekely
Forschungszentrum Jülich GmbH, JCNS @ MLZ , Lichtenbergstraße 1, 85747 Garching, Germany.
Vitaliy
V
Pipich
European Synchrotron Radiation Facility , 71 avenue des Martyrs, 38043 Grenoble Cedex 9, France.
Michael
M
Sztucki
Institut für Angewandte Physik, Universität Tübingen , Auf der Morgenstelle 10, 72076 Tübingen, Germany.
Frank
F
Schreiber
D003460:Crystallization; D007782:Lactoglobulins / Q000737:chemistry*; D033363:Neutron Diffraction; D066329:Protein Aggregates / Q000502:physiology*; D053838:Scattering, Small Angle; D017550:Spectroscopy, Fourier Transform Infrared; D014961:X-Ray Diffraction; D015019:Yttrium / Q000737:chemistry*
D007782:Lactoglobulins; D066329:Protein Aggregates; C064782:yttrium chloride; D015019:Yttrium