Protein cluster formation in aqueous solution in the presence of multivalent metal ions--a light scattering study.
The formation of protein clusters as precursors for crystallization and phase separation is of fundamental and practical interest in protein science. Using multivalent ions, the strengths of both long-range Coulomb repulsion and short-range attraction can be tuned in protein solutions, representing a well-controlled model system to study static and dynamic properties of clustering during the transition from a charge-stabilized to an aggregate regime. Here, we study compressibility, diffusion, and size of solutes by means of static (SLS) and dynamic light scattering (DLS) in solutions of bovine serum albumin (BSA) and YCl3. For this and comparable systems, an increasing screening and ultimately inversion of the protein surface charge induce a rich phase behavior including reentrant condensation, liquid-liquid phase separation and crystallization, which puts the cluster formation in the context of precursor formation and nucleation of liquid and crystalline phases. We find that, approaching the turbid aggregate regime with increasing salt concentration cs, the diffusion coefficients decrease and the scattered intensity increases by orders of magnitude, evidencing increasing correlation lengths likely associated with clustering. The combination of static and dynamic observations suggests the formation of BSA clusters with a size on the order of 100 nm. The global thermodynamic state seems to be stable over at least several hours. Surprisingly, results on collective diffusion and inverse compressibility from different protein concentrations can be rescaled into master curves as a function of cs/c*, where c* is the critical salt concentration of the transition to the turbid aggregate regime.
Institut für Angewandte Physik, Universität Tübingen, Auf der Morgenstelle 10, 72076 Tübingen, Germany. fajun.zhang@uni-tuebingen.de.
Daniel
D
Soraruf
Felix
F
Roosen-Runge
Marco
M
Grimaldo
Fabio
F
Zanini
Ralf
R
Schweins
Tilo
T
Seydel
Fajun
F
Zhang
Roland
R
Roth
Martin
M
Oettel
Frank
F
Schreiber
D000818:Animals; D002417:Cattle; D003460:Crystallization; D007477:Ions / Q000737:chemistry*; D012542:Scattering, Radiation; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D014867:Water / Q000737:chemistry
D007477:Ions; D012996:Solutions; D014867:Water; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride