β-Crystallins of the adult chicken lens

Relatedness of the polypeptides and their aggregates

Harry Ostrer, Joram Piatigorsky

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

β-Crystallins, the major class of polymeric soluble proteins in the mature lens, were isolated from one-year-old chickens by Sephadex G200 chromatography and identified by immunodiffusion. The β-crystallin polypeptides were resolved into six components with molecular weights near 35 K, 29 K, 26 K, 24 K, 20 K and 19 K by SDS-urea-polyacrylamide gel electrophoresis. The β-crystallins eluting later from the column contained relatively less of the 35 K polypeptides. Two-dimensional tryptic fingerprints of iodinated peptides indicated that the β-crystallin polypeptides had related primary structures, although some differences in the peptide maps were evident. Isoelectric focusing of undissociated β-crystallins in a sucrose density gradient resolved four major forms with isoelectric points of 4·6, 6·0, 6·4 and 6·6. Preparative isoelectric focusing proved to be an effective method for purification of these proteins. SDS-urea-polyacrylamide gel electrophoresis showed that each of the major isoelectric forms of β-crystallin had a characteristic combination of polypeptides. The 35 K and 29 K polypeptides were present only in the most acidic β-crystallins. Finally, circular dichroism spectroscopy showed that, despite their different polypeptide compositions, the four β-crystallins had similar secondary and tertiary conformations.

Original languageEnglish (US)
Pages (from-to)679-689
Number of pages11
JournalExperimental Eye Research
Volume30
Issue number6
DOIs
StatePublished - 1980

Fingerprint

Crystallins
Lenses
Chickens
Peptides
Isoelectric Focusing
Urea
Polyacrylamide Gel Electrophoresis
Peptide Mapping
Immunodiffusion
Isoelectric Point
Circular Dichroism
Sucrose
Chromatography
Spectrum Analysis
Proteins
Molecular Weight

Keywords

  • chicken
  • circular dichroism
  • crystallin
  • isoelectric focusing
  • lens
  • polyacrylamide gel electrophoresis
  • radioiodination
  • tryptic peptide maps

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

β-Crystallins of the adult chicken lens : Relatedness of the polypeptides and their aggregates. / Ostrer, Harry; Piatigorsky, Joram.

In: Experimental Eye Research, Vol. 30, No. 6, 1980, p. 679-689.

Research output: Contribution to journalArticle

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