Characterization of γ-Crystallins from a Hybrid Teleostean Fish

Multiplicity of Isoforms as Revealed by cDNA Sequence Analysis

F. M. Pan, W. C. Chang, Y. K. Chao, S. H. Chiou

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

γ-Crystallins were isolated and characterized from the eye lenses of a hybrid species belonging to the teleostean fish. Isoelectric focusing of γ-crystallin fraction obtained from gel-permeation chromatography revealed that it consists of multiple charge isomers of a protein species with a molecular mass of about 20 kDa. To facilitate the cloning of γ-crystallin gene, cDNA was constructed from the poly(A)+mRNA isolated from fresh lenses, and amplification by polymerase chain reaction (PCR) was carried out to obtain cDNA encoding multiple γ-crystallins. Sequencing five of more than 10 positive clones revealed that a multiplicity of isoforms exists in the γ-crystallin class of teleostean lenses. Comparison of protein sequences encoded by these multiple cDNAs with those published sequences of γ-crystallins from bovine, mouse and carp lenses indicated that there is about 70-80 % sequence homology between different species of piscine species whereas only 50-60 % is found between mammals and fishes. Structural analysis of these γ-crystallins with high methionine contents (11-16 %) suggests that there are two major subclasses of piscine γ-crystallins, i.e. γM1 and γM2, existed long before the appearance of mammalian γ-crystallin with low methionines.

Original languageEnglish
Pages (from-to)527-534
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume202
Issue number1
DOIs
Publication statusPublished - Jul 15 1994
Externally publishedYes

Fingerprint

Crystallins
Fish
Sequence Analysis
Protein Isoforms
Fishes
Complementary DNA
Lenses
Methionine
Crystalline Lens
Mammals
Carps
Cloning
Polymerase chain reaction
Gel permeation chromatography
Isoelectric Focusing
Molecular mass
Sequence Homology
Structural analysis
Isomers
Gel Chromatography

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Characterization of γ-Crystallins from a Hybrid Teleostean Fish : Multiplicity of Isoforms as Revealed by cDNA Sequence Analysis. / Pan, F. M.; Chang, W. C.; Chao, Y. K.; Chiou, S. H.

In: Biochemical and Biophysical Research Communications, Vol. 202, No. 1, 15.07.1994, p. 527-534.

Research output: Contribution to journalArticle

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