April 1964
Volume 3, Issue 2
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Articles  |   April 1964
Methods of Isolation of Alpha, Beta, and Gamma Crystallins and their Subgroups
Author Affiliations
  • ABRAHAM SPECTOR
    Howe Laboratory of Ophthalmology, Harvard Medical School, and the Massachusetts Eye and Ear Infirmary, Boston, Mass.
Investigative Ophthalmology & Visual Science April 1964, Vol.3, 182-193. doi:
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      ABRAHAM SPECTOR; Methods of Isolation of Alpha, Beta, and Gamma Crystallins and their Subgroups. Invest. Ophthalmol. Vis. Sci. 1964;3(2):182-193.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

It is now possible to separate alpha, beta, and gamma crystallins by a relatively simple procedure combining precipitation with zinc glycinate, and isoelectric precipitation followed by sephadex 75 chromatography. Paper electrophoresis and ultracentrifugal analyses indicate that the isolated crystallins are similar to material isolated by other procedures and contain only minor contaminants.

Calf lens protein components separated by DEAE chromatography have been related to the crystallins isolated by the new method. Gamma crystallin is the first fraction eluted from the column, followed by the beta crystallin components, and then the alpha crystallin subgroup. Since the DEAE separable components could thus be identified as members of the alpha, beta, or gamma crystallin subgroups, another method is provided for isolating the crystallins.

The properties of the crystallins obtained by the two methods were studied. Electrophoretic and ultracentrifugal analyses indicate that the alpha and gamma fractions isolated by DEAE chromatography are similar to crystallins isolated by the zinc glycinate procedure. However, the beta crystallin fractions isolated by the two procedures appear to have similar electrophoretic mobilities but differ in their sedimentation properties.

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