Cited by 93Enzyme Structure and FunctionBiochemical and Molecular ResearchMetal-Catalyzed Oxygenation Mechanisms

The Conversion from the Dehydrogenase Type to the Oxidase Type of Rat Liver Xanthine Dehydrogenase by Modification of Cysteine Residues with Fluorodinitrobenzene

Open access
Nishino, Tomoko, Nishino, Takeshi·1997·Journal of Biological Chemistry

Abstract

When rat liver xanthine dehydrogenase was incubated with fluorodinitrobenzene (FDNB) at pH 8.5, the total enzyme activity decreased gradually to a limited value of initial activity with modification of two lysine residues in a similar way to the modification of bovine milk xanthine oxidase with FDNB (Nishino, T., Tsushima, K., Hille, R. and Massey, V. (1982) J. Biol. Chem. 257, 7348-7353). After modification with FDNB, the two peptides containing dinitrophenyl-lysine were isolated from the molybdopterin domain after proteolytic digestion and were identified as Lys754 and Lys771 by sequencing the peptides. During the modification of these lysine residues, xanthine dehydrogenase was found to be converted to an oxidase form in the early stage of incubation. Incorporation of the 3H-dinitrophenyl group into enzyme cysteine residues was 0.96 mol per enzyme FAD for 68% conversion to the oxidase form. The modified enzyme was reconverted to the dehydrogenase form by incubation with dithiothreitol with concomitant release of 3H-dinitrophenyl compounds. After modification with 3H-FDNB followed by carboxymethylation under denaturating conditions, the enzyme was digested with proteases. Three 3H-dinitrophenyl-labeled peptides were isolated and sequenced. The modified residues were identified to be Cys535, Cys992 and Cys1324. These residues are conserved among the all known mammalian enzymes, but Cys992 and Cys1324 are not conserved in the chicken enzyme. Cys1324 of the rat enzyme was found not to be involved in the conversion from the dehydrogenase to the oxidase by limited proteolysis experiments, but Cys535 and Cys992 which seemed to be modified alternatively with FDNB appear to be involved in the conversion.

Integrity signals

Registry

Crossref · 10.1074/jbc.272.47.29859

Cites

34 of 34 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

We couldn’t retrieve this paper’s full text, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Authors
Nishino, T., & Nishino, T.
Affiliations
Nippon Medical School (JP)
Corresponding author
Nishino, Tomoko
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1997-11
Volume
272
Issue
47
Pages
29859-29864
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2009887805
Cited by
93
References
34
Citations by year
2024: 2 · 2023: 5 · 2022: 1 · 2021: 2 · 2020: 1 · 2019: 4 · 2018: 2 · 2017: 2 · 2016: 2 · 2015: 5 · 2014: 6 · 2013: 2 · 2012: 8
FWCI
1.63
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
  • Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
Keywords
xanthine dehydrogenase, xanthine oxidase, fluorodinitrobenzene, cysteine modification, lysine modification, rat liver enzyme
MeSH terms
Amino Acid Sequence, Animals, Cattle, Chickens, Chromatography, High Pressure Liquid, Cysteine, Dinitrofluorobenzene, Liver, Lysine, Molecular Sequence Data, Spectrophotometry, Atomic, Xanthine Dehydrogenase, Xanthine Oxidase, Rats

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review