TY - JOUR
T1 - λ-crystallin related to dehydroascorbate reductase in the rabbit lens
AU - Suzuki, Takahiro
AU - Bando, Masayasu
AU - Oka, Mikako
AU - Tsukamoto, Hideo
AU - Akatsuka, Ichiko
AU - Kawai, Kenji
AU - Obazawa, Hajime
AU - Kobayashi, Shizuko
AU - Takehana, Makoto
PY - 2003/1/1
Y1 - 2003/1/1
N2 - 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.
AB - 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.
KW - Dehydroascorbate reductase
KW - Enzyme-crystallin
KW - Heterogeneity
KW - Rabbit lens
KW - λ-Crystallin
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U2 - 10.1016/S0021-5155(03)00140-0
DO - 10.1016/S0021-5155(03)00140-0
M3 - Article
C2 - 12967857
AN - SCOPUS:0041335144
VL - 47
SP - 437
EP - 443
JO - Japanese Journal of Ophthalmology
JF - Japanese Journal of Ophthalmology
SN - 0021-5155
IS - 5
ER -