λ-crystallin related to dehydroascorbate reductase in the rabbit lens

Takahiro Suzuki, Masayasu Bando, Mikako Oka, Hideo Tsukamoto, Ichiko Akatsuka, Kenji Kawai, Hajime Obazawa, Shizuko Kobayashi, Makoto Takehana

研究成果: Article査読

5 被引用数 (Scopus)

抄録

Purpose: To evaluate the relationship of λ-crystallin to reduced nicotinamide adenine dinucleotide (NADH)-dependent dehydroascorbate (DHA) reductase found specifically in the rabbit lens. Methods: DHA reductase Fractions I-IV were separated from the λ/βL1-crystallin fraction of rabbit lens soluble protein by diethylaminoethyl (DEAE)-cellulose ion-exchange column chromatography, and then the enzyme was partially purified from Fraction II by rechromatography on the same ion-exchange column. The isolated DHA reductase fractions were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting, native isoelectric focusing and two-dimensional gel electrophoresis. Results: Using Western blot and a probe of antiserum to recombinant λ-crystallin, the main 33-kDa protein band was strongly stained in all the rabbit lens DHA reductase fractions, and most of the additional protein bands of approximately 25-30 kDa were also detectable. In the partially purified enzyme, the 33-kDa subunit alone was identified as a distinct protein band by SDS-PAGE, and a main basic protein was found at pI 7.6 by native isoelectric focusing. In addition, many bands of more acidic proteins were separated from other enzyme fractions, and protein spots corresponding to the 33 and/or approximately 25-30-kDa subunits were detected in each of the more acidic proteins by two-dimensional gel electrophoresis. Conclusion: These results suggest that λ-crystallin is closely related to the DHA reductase in the rabbit lens. The above heterogeneity of the enzyme-crystallin may arise from posttranslational modifications.

本文言語English
ページ(範囲)437-443
ページ数7
ジャーナルJapanese Journal of Ophthalmology
47
5
DOI
出版ステータスPublished - 2003

ASJC Scopus subject areas

  • 眼科学

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