On the equilibrium between monomeric alpha-lactalbumin and the chaperoning complex of alpha-crystallin.
In chaperoning dithiothreitol-denatured alpha-lactabumin, alpha-crystallin forms a chaperoning complex. In order to study the kinetics of such chaperoning it needs to be established whether the formation of the chaperoning complex is a reversible or irreversible process. The chaperoning reaction was studied by dynamic light scattering as a function of concentration and weight ratio of alpha-lactalbumin/alpha-crystallin. HPLC and subsequent SDS-PAGE gel electrophoresis experiments established that the chaperoning complex formed contains both alpha-crystallin and alpha-lactalbumin. Upon rechromatographing the chaperoning complex, the presence of monomeric alpha-lactalbumin has been demonstrated in addition to the chaperoning complex itself. This and equilibrium dialysis experiments demonstrated conclusively the existence of an equilibrium between monomeric partially denatured alpha-lactalbumin and the chaperoning complex made of alpha-lactalbumin and alpha-crystallin.
Neal
R
R
Laboratory of Mechanisms of Ocular Disease, National Eye Institute, Bethesda, Maryland 20892-2735, USA.
Zigler
J S
JS
Bettelheim
F A
FA
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000737:chemistry*; D004229:Dithiothreitol; D004591:Electrophoresis, Polyacrylamide Gel; D007768:Lactalbumin / Q000737:chemistry*; D007908:Lens, Crystalline / Q000737:chemistry; D008027:Light; D018832:Molecular Chaperones / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry; D011489:Protein Denaturation; D012542:Scattering, Radiation
D003459:Crystallins; D018832:Molecular Chaperones; D007768:Lactalbumin; D009113:Muramidase; D004229:Dithiothreitol