Further Studies on myo-Inositol-1-phosphatase from the Pollen of Lilium longiflorum Thunb.
PDF (1.1M)
Journal: 2010/July - Plant Physiology
ISSN: 0032-0889
PUBMED: 16663819
Abstract:
myo-Inositol-1-phosphatase has been purified to homogeneity from Lilium longiflorum pollen using an alternative procedure which includes pH change and phenyl Sepharose column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis shows that the enzyme is a dimer (subunit molecular weight, 29,000 daltons). The enzyme is stable at low pH values and is inactivated only below pH 3.0. In addition to 1l-and 1d-myo-inositol-1-phosphate, it shows high specificity for 1l-chiro-inositol-3-phosphate. As observed earlier with other primary phosphate esters, d-glucitol-6-phosphate and d-mannitol-6-phosphate are hydrolyzed very slowly. No activity is observed with inorganic pyrophosphate or myo-inositol pentaphosphate as substrate. The enzyme is inhibited by fluoride, sulfate, molybdate, and thiol-directed reagents. Partial protection against N-ethylmaleimide inhibition by substrate and Mg(2+) together suggests sulfhydryl involvement at the active site.
Open in
PUBMED | PMC | Google Scholar | Wikipedia
Relations:
Content
Citations
(14)
References
(13)
Drugs
(3)
Affiliates
(1)
Similar articles
Articles by the same authors
Discussion board
Plant Physiol 76(1): 40-44
PMID: 16663819

Further Studies on <em>myo</em>-Inositol-1-phosphatase from the Pollen of <em>Lilium longiflorum</em> Thunb <sup><a href="#fn1" rid="fn1" class=" fn">1</a></sup>

Abstract

myo-Inositol-1-phosphatase has been purified to homogeneity from Lilium longiflorum pollen using an alternative procedure which includes pH change and phenyl Sepharose column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis shows that the enzyme is a dimer (subunit molecular weight, 29,000 daltons). The enzyme is stable at low pH values and is inactivated only below pH 3.0. In addition to 1l-and 1d-myo-inositol-1-phosphate, it shows high specificity for 1l-chiro-inositol-3-phosphate. As observed earlier with other primary phosphate esters, d-glucitol-6-phosphate and d-mannitol-6-phosphate are hydrolyzed very slowly. No activity is observed with inorganic pyrophosphate or myo-inositol pentaphosphate as substrate. The enzyme is inhibited by fluoride, sulfate, molybdate, and thiol-directed reagents. Partial protection against N-ethylmaleimide inhibition by substrate and Mg together suggests sulfhydryl involvement at the active site.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
icon of scanned page 40
40
icon of scanned page 41
41
icon of scanned page 42
42
icon of scanned page 43
43
icon of scanned page 44
44

Images in this article

Fig. 4
Fig. 4
on p.42
Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Baykov AA, Artjukov AA, Avaeva SM. Fluoride inhibition of inorganic pyrophosphatase. I. Kinetic studies in a Mg2+-PPi system using a new continuous enzyme assay. Biochim Biophys Acta. 1976 May 13;429(3):982–992. [PubMed] [Google Scholar]
  • Berridge MJ. Rapid accumulation of inositol trisphosphate reveals that agonists hydrolyse polyphosphoinositides instead of phosphatidylinositol. Biochem J. 1983 Jun 15;212(3):849–858.[PMC free article] [PubMed] [Google Scholar]
  • Blakesley RW, Boezi JA. A new staining technique for proteins in polyacrylamide gels using coomassie brilliant blue G250. Anal Biochem. 1977 Oct;82(2):580–582. [PubMed] [Google Scholar]
  • Dunker AK, Rueckert RR. Observations on molecular weight determinations on polyacrylamide gel. J Biol Chem. 1969 Sep 25;244(18):5074–5080. [PubMed] [Google Scholar]
  • Eisenberg F., Jr D-myoinositol 1-phosphate as product of cyclization of glucose 6-phosphate and substrate for a specific phosphatase in rat testis. J Biol Chem. 1967 Apr 10;242(7):1375–1382. [PubMed] [Google Scholar]
  • DE GUBAREFF T, FURCHGOTT RF. The determination of inorganic phosphate and creatine phosphate in tissue extracts. J Biol Chem. 1956 Nov;223(1):377–388. [PubMed] [Google Scholar]
  • Hallcher LM, Sherman WR. The effects of lithium ion and other agents on the activity of myo-inositol-1-phosphatase from bovine brain. J Biol Chem. 1980 Nov 25;255(22):10896–10901. [PubMed] [Google Scholar]
  • Joseph SK, Thomas AP, Williams RJ, Irvine RF, Williamson JR. myo-Inositol 1,4,5-trisphosphate. A second messenger for the hormonal mobilization of intracellular Ca2+ in liver. J Biol Chem. 1984 Mar 10;259(5):3077–3081. [PubMed] [Google Scholar]
  • King J, Laemmli UK. Polypeptides of the tail fibres of bacteriophage T4. J Mol Biol. 1971 Dec 28;62(3):465–477. [PubMed] [Google Scholar]
  • Loewus MW, Loewus FA. myo-Inositol-1-Phosphatase from the Pollen of Lilium longiflorum Thunb. Plant Physiol. 1982 Sep;70(3):765–770.[PMC free article] [PubMed] [Google Scholar]
  • Naccarato WF, Ray RE, Wells WW. Biosynthesis of myo-inositol in rat mammary gland. Isolation and properties of the enzymes. Arch Biochem Biophys. 1974 Sep;164(1):194–201. [PubMed] [Google Scholar]
  • Roth SC, Harkness DR, Isaacks RE. Studies on avian erythrocyte metabolism: purification and properties of myo-inositol 1-phosphatase form chick erythrocytes. Arch Biochem Biophys. 1981 Sep;210(2):465–473. [PubMed] [Google Scholar]
  • Sedmak JJ, Grossberg SE. A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem. 1977 May 1;79(1-2):544–552. [PubMed] [Google Scholar]
Program in Biochemistry/Biophysics and the Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340
Supported in part by Grant GM-22427 from the National Institute of Health, United States Public Health Service. Scientific Paper No. 6763, Project 0266, College of Agriculture Research Center, Washington State University, Pullman, WA 99164.
Copyright notice
Abstract
myo-Inositol-1-phosphatase has been purified to homogeneity from Lilium longiflorum pollen using an alternative procedure which includes pH change and phenyl Sepharose column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis shows that the enzyme is a dimer (subunit molecular weight, 29,000 daltons). The enzyme is stable at low pH values and is inactivated only below pH 3.0. In addition to 1l-and 1d-myo-inositol-1-phosphate, it shows high specificity for 1l-chiro-inositol-3-phosphate. As observed earlier with other primary phosphate esters, d-glucitol-6-phosphate and d-mannitol-6-phosphate are hydrolyzed very slowly. No activity is observed with inorganic pyrophosphate or myo-inositol pentaphosphate as substrate. The enzyme is inhibited by fluoride, sulfate, molybdate, and thiol-directed reagents. Partial protection against N-ethylmaleimide inhibition by substrate and Mg together suggests sulfhydryl involvement at the active site.
Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.