Biophysical characterization of alpha-crystallin aggregates: validation of the micelle hypothesis.
The size of alpha-crystallin aggregates, as well as the structural organization of each particle's subunits, is currently unknown, although a number of different laboratories have suggested both structures and average molecular weights (Thomson, J.A. and Augusteyn, R.C. (1984) Proc. Int. Soc. Eye Res. 3, 152). One hypothesis, compatible with literature reports and consistent with what is known of subunit primary structure and physiological function, is that alpha-crystallin exists in vivo as a naturally occurring protein micelle (Sen, A.C. and Chakrabarti, B. (1991) Biophysical J. 59, 108a.) To test this hypothesis, experiments were performed on this protein to determine its behavior under increased hydrostatic pressure and the effect of its concentration on aqueous surface tension. With increasing hydrostatic pressure, the turbidity of an alpha-crystallin solution increases exponentially to a plateau at about 6000-8000 psi; upon release of pressure, the samples slowly return to their original turbidity level. Other naturally aggregating proteins, such as skeletal muscle myosin, demonstrate a decrease in turbidity under the same conditions. The surface tension of alpha-crystallin in aqueous solution decreases to a plateau with increasing protein subunit concentration, with an inflection point over the range 0.18-0.25 mM; cholate and other amphiphiles exhibit similar behavior. In contrast, plots of surface tension over the equivalent concentration range for other protein aggregates in the same buffer more closely approximate the types of curve obtained with short chain aliphatic acids. These results indicate that alpha-crystallin behaves like the protein version of a micelle.
Radlick
L W
LW
Center for Biophysics, Rensselaer Polytechnic Institute, Troy, NY 12180-3590.
Koretz
J F
JF
D000818:Animals; D002417:Cattle; D003459:Crystallins / Q000737:chemistry*; D006874:Hydrostatic Pressure; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008823:Micelles; D008854:Microscopy, Electron; D009391:Nephelometry and Turbidimetry
D003459:Crystallins; D046911:Macromolecular Substances; D008823:Micelles