RRC ID 19854
著者 Kang CB, Ho KK.
タイトル Characterization of a soluble inorganic pyrophosphatase from Microcystis aeruginosa and preparation of its antibody.
ジャーナル Arch Biochem Biophys
Abstract A soluble inorganic pyrophosphatase was isolated from a crude extract of Microcystis aeruginosa by adsorption chromatography. The enzyme was purified to homogeneity as judged by sodium dodecyl sulfate (SDS) and nondenaturing polyacrylamide gel electrophoresis and N-terminal amino acid analysis. The molecular mass was estimated to be 80 kDa by gel filtration chromatography, 87 kDa by nondenaturing polyacrylamide gel electrophoresis, and 28 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme has an isoelectric point of 4.5, which is similar to the pI values reported for other soluble inorganic pyrophosphatases. The sequence of 29 N-terminal amino acids was determined; only 4 of these amino acids are identical to those in the sequence of Saccharomyces cerevisiae inorganic pyrophosphatase. M. aeruginosa inorganic pyrophosphatase is a Mg(2+)-dependent enzyme exhibiting a pH optimum of around 7.5. Its KM value for inorganic pyrophosphate was estimated to be 1.30 mM. A specific antibody was raised in chicken to M. aeruginosa inorganic pyrophosphatase. No immunological cross-reactivity was seen when Western blots of partially purified S. cerevisiae or Escherichia coli inorganic pyrophosphatase were probed with the antibody.
巻・号 289(2)
ページ 281-8
公開日 1991-9-1
DOI 10.1016/0003-9861(91)90473-v
PII 0003-9861(91)90473-V
PMID 1654852
MeSH Amino Acid Sequence Animals Antibodies Inorganic Pyrophosphatase Kinetics Microcystis / enzymology* Molecular Sequence Data Molecular Weight Protein Conformation Pyrophosphatases / immunology Pyrophosphatases / isolation & purification* Pyrophosphatases / metabolism Solubility
IF 3.391
引用数 8
WOS 分野 BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
藻類 NIES-44