Oligomerization and phase transitions in aqueous solutions of native and truncated human beta B1-crystallin.
Human betaB1-crystallin is a major eye-lens protein that undergoes in vivo truncation at the N-terminus with aging. By studying native betaB1 and truncated betaB1DeltaN41, which mimics an age-related in vivo truncation, we have determined quantitatively the effect of truncation on the oligomerization and phase transition properties of betaB1 aqueous solutions. The oligomerization studies show that the energy of attraction between the betaB1DeltaN41 proteins is about 10% greater than that of the betaB1 proteins. We have found that betaB1DeltaN41 aqueous solutions undergo two distinct types of phase transitions. The first phase transition involves an initial formation of thin rodlike assemblies, which then evolve to form crystals. The induction time for the formation of rodlike assemblies is sensitive to oligomerization. The second phase transition can be described as liquid-liquid phase separation (LLPS) accompanied by gelation within the protein-rich phase. We refer to this process as heterogeneous gelation. These two phase transitions are not observed in the case of betaB1 aqueous solutions. However, upon the addition of poly(ethylene glycol) (PEG), we observe heterogeneous gelation also for betaB1. Our PEG experiments allow us to estimate the difference in phase separation temperatures between betaB1 and betaB1DeltaN41. This difference is consistent with the increase in energy of attraction found in our oligomerization studies. Our work suggests that truncation is a cataractogenic modification since it favors protein condensation and the consequent formation of light scattering elements, and highlights the importance of the N-terminus of betaB1 in maintaining lens transparency.
Annunziata
Onofrio
O
Department of Physics, Center for Materials Science and Engineering, and Materials Processing Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139-4307, USA.
Pande
Ajay
A
Pande
Jayanti
J
Ogun
Olutayo
O
Lubsen
Nicolette H
NH
Benedek
George B
GB
D002942:Circular Dichroism; D003459:Crystallins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008027:Light; D044367:Phase Transition; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012542:Scattering, Radiation; D017384:Sequence Deletion; D012996:Solutions; D013816:Thermodynamics; D014867:Water / Q000737:chemistry; D038206:beta-Crystallin B Chain / Q000737:chemistry*
C105601:CRYBB1 protein, human; D003459:Crystallins; D011994:Recombinant Proteins; D012996:Solutions; D038206:beta-Crystallin B Chain; D014867:Water