| RRC ID |
19854
|
| Author |
Kang CB, Ho KK.
|
| Title |
Characterization of a soluble inorganic pyrophosphatase from Microcystis aeruginosa and preparation of its antibody.
|
| Journal |
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.
|
| Volume |
289(2)
|
| Pages |
281-8
|
| Published |
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
|
| Times Cited |
8
|
|
WOS Category
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Algae |
NIES-44 |
