Intermolecular exchange and stabilization of recombinant human alphaA- and alphaB-crystallin

J Biol Chem. 1998 Jan 2;273(1):286-90. doi: 10.1074/jbc.273.1.286.

Abstract

Lens alpha-crystallin subunits alphaA and alphaB are differentially expressed and have a 3-to-1 ratio in most mammalian lenses by intermolecular exchange. The biological significance of this composition and the mechanism of exchange are not clear. Preparations of human recombinant alphaA- and alphaB-crystallins provide a good system in which to study this phenomenon. Both recombinant alphaA- and alphaB-crystallins are folded and aggregated to the size of the native alpha-crystallin. During incubation together, they undergo an intermolecular exchange as shown by native isoelectric focusing. Circular dichroism measurements indicate that the protein with a 3-to-1 ratio of alphaA- and alphaB-crystallins has the same secondary structure but somewhat different tertiary structures after exchange: the near-UV CD increases after exchange. The resulting hybrid aggregate is more stable than the individual homogeneous aggregates: at 62 degrees C, alphaB-crystallin is more susceptible to aggregation and displays a greater light scattering than alphaA-crystallin. This heat-induced aggregation of alphaB-crystallin, however, was suppressed by intermolecular exchange with alphaA-crystallin. These phenomena are also observed by fast performance liquid chromatography gel filtration patterns. The protein structure of alphaB-crystallin is stabilized by intermolecular exchange with alphaA-crystallin.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Chromatography, Gel
  • Chromatography, Liquid
  • Circular Dichroism
  • Crystallins / chemistry*
  • Humans
  • Infant
  • Light
  • Recombinant Proteins / chemistry
  • Scattering, Radiation

Substances

  • Crystallins
  • Recombinant Proteins