TY - JOUR
T1 - Isolation of ascorbate free radical reductase from rabbit lens soluble fraction
AU - Bando, Masayasu
AU - Inoue, Takashi
AU - Oka, Mikako
AU - Nakamura, Kayako
AU - Kawai, Kenji
AU - Obazawa, Hajime
AU - Kobayashi, Shizuko
AU - Takehana, Makoto
PY - 2004/12
Y1 - 2004/12
N2 - Ascorbate free radical (AFR) reductase with diaphorase activity was isolated from the rabbit lens soluble fraction to characterise some molecular properties of the enzyme. The isolation was accomplished using gel filtration (Sephadex G-75 superfine or Sephacryl S-200 HR), affinity chromatography (Affi-Gel Blue), native isoelectric focusing and two-dimensional gel electrophoresis. A major soluble AFR reductase was found at an isoelectric point of 8·4 and a molecular weight of 31 kDa, and a few minor enzymes were also detected in the range of pI 7·0-8·6. An unknown N-terminal partial amino acid sequence was determined in one peptide fragment prepared from the major enzyme fraction. From the sequence analysis, it is discussed that the lens soluble AFR reductase may differ from NADH-cytochrome b 5 reductase reported to be involved in the membrane-bound AFR reductase activity of mitochondria, microsomes and plasma membrane.
AB - Ascorbate free radical (AFR) reductase with diaphorase activity was isolated from the rabbit lens soluble fraction to characterise some molecular properties of the enzyme. The isolation was accomplished using gel filtration (Sephadex G-75 superfine or Sephacryl S-200 HR), affinity chromatography (Affi-Gel Blue), native isoelectric focusing and two-dimensional gel electrophoresis. A major soluble AFR reductase was found at an isoelectric point of 8·4 and a molecular weight of 31 kDa, and a few minor enzymes were also detected in the range of pI 7·0-8·6. An unknown N-terminal partial amino acid sequence was determined in one peptide fragment prepared from the major enzyme fraction. From the sequence analysis, it is discussed that the lens soluble AFR reductase may differ from NADH-cytochrome b 5 reductase reported to be involved in the membrane-bound AFR reductase activity of mitochondria, microsomes and plasma membrane.
KW - ascorbate free radical reductase
KW - isolation
KW - partial amino acid sequence
KW - rabbit lens
KW - soluble fraction
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U2 - 10.1016/j.exer.2004.08.011
DO - 10.1016/j.exer.2004.08.011
M3 - Article
C2 - 15642324
AN - SCOPUS:11844302306
SN - 0014-4835
VL - 79
SP - 869
EP - 873
JO - Experimental Eye Research
JF - Experimental Eye Research
IS - 6
ER -