Two-dimensional gel electrophoretic analysis of human Lens proteins

Manuel B. Datiles, D. James Schumer, J. Samuel Zigler, Paul Russell, Leigh Anderson, Donita Garland

Research output: Contribution to journalArticle

Abstract

Human lens proteins from clear lenses were separated and identified using two-dimensional polyacrylamide electrophoresis. Isoelectric focusing, both equilibrium and non-equilibrium, was performed in the first dimension and SDS electrophoresis in the second dimension. Proteins were identified by Western blotting and sequencing techniques and by comparison with patterns obtained with purified crystallin fractions. Analyses were performed on total urea soluble proteins of lenses varying in age from fetal to 73 yr. Several hundred protein spots representing crystallins, cytoskeletal proteins and enzymes were resolved in the fetal lens. In the older lenses there was a dramatic increase in the number of protein species in the molecular weight range of the crystallins and a reduced number of discrete protein species visible at molecular weights greater than 50,000. Conversely, a number of proteins below approximately 15 kDa were visible even in the fetal lens. The number and amount of polypeptides in this molecular weight range were increased in the older lenses. Many of these low molecular weight species could be assigned to either the α β or γcrystallin fractions. An age dependent increase in the number of acidic species of both crystallins and other proteins, such as, glyceraldehyde 3-phosphate dehydrogenase was observed as well as the loss or mobility change of γcrystallin. Two-dimensional gel electrophoresis provides a sensitive and practical technique for characterizing all of the proteins of the human lens.

Original languageEnglish (US)
Pages (from-to)669-677
Number of pages9
JournalCurrent Eye Research
Volume11
Issue number7
DOIs
StatePublished - Jan 1 1992

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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    Datiles, M. B., Schumer, D. J., Zigler, J. S., Russell, P., Anderson, L., & Garland, D. (1992). Two-dimensional gel electrophoretic analysis of human Lens proteins. Current Eye Research, 11(7), 669-677. https://doi.org/10.3109/02713689209000740