Isolation of ascorbate free radical reductase from rabbit lens soluble fraction

Masayasu Bando, Takashi Inoue, Mikako Oka, Kayako Nakamura, Kenji Kawai, Hajime Obazawa, Shizuko Kobayashi, Makoto Takehana

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)869-873
Number of pages5
JournalExperimental Eye Research
Volume79
Issue number6
DOIs
Publication statusPublished - 2004 Dec

Fingerprint

Lenses
Rabbits
Cibacron Blue F 3GA
Enzymes
Cytochromes b5
Peptide Fragments
Electrophoresis, Gel, Two-Dimensional
Isoelectric Point
Isoelectric Focusing
Microsomes
Affinity Chromatography
NAD
Gel Chromatography
Sequence Analysis
Amino Acid Sequence
Mitochondria
Oxidoreductases
Molecular Weight
Cell Membrane
Membranes

Keywords

  • ascorbate free radical reductase
  • isolation
  • partial amino acid sequence
  • rabbit lens
  • soluble fraction

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Bando, M., Inoue, T., Oka, M., Nakamura, K., Kawai, K., Obazawa, H., ... Takehana, M. (2004). Isolation of ascorbate free radical reductase from rabbit lens soluble fraction. Experimental Eye Research, 79(6), 869-873. https://doi.org/10.1016/j.exer.2004.08.011

Isolation of ascorbate free radical reductase from rabbit lens soluble fraction. / Bando, Masayasu; Inoue, Takashi; Oka, Mikako; Nakamura, Kayako; Kawai, Kenji; Obazawa, Hajime; Kobayashi, Shizuko; Takehana, Makoto.

In: Experimental Eye Research, Vol. 79, No. 6, 12.2004, p. 869-873.

Research output: Contribution to journalArticle

Bando, M, Inoue, T, Oka, M, Nakamura, K, Kawai, K, Obazawa, H, Kobayashi, S & Takehana, M 2004, 'Isolation of ascorbate free radical reductase from rabbit lens soluble fraction', Experimental Eye Research, vol. 79, no. 6, pp. 869-873. https://doi.org/10.1016/j.exer.2004.08.011
Bando, Masayasu ; Inoue, Takashi ; Oka, Mikako ; Nakamura, Kayako ; Kawai, Kenji ; Obazawa, Hajime ; Kobayashi, Shizuko ; Takehana, Makoto. / Isolation of ascorbate free radical reductase from rabbit lens soluble fraction. In: Experimental Eye Research. 2004 ; Vol. 79, No. 6. pp. 869-873.
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