Purification and properties of human placental ATP diphosphohydrolase
| dc.contributor.author | Christoforidis, S. | en |
| dc.contributor.author | Papamarcaki, T. | en |
| dc.contributor.author | Galaris, D. | en |
| dc.contributor.author | Kellner, R. | en |
| dc.contributor.author | Tsolas, O. | en |
| dc.date.accessioned | 2015-11-24T18:55:28Z | |
| dc.date.available | 2015-11-24T18:55:28Z | |
| dc.identifier.issn | 0014-2956 | - |
| dc.identifier.uri | https://olympias.lib.uoi.gr/jspui/handle/123456789/18885 | |
| dc.rights | Default Licence | - |
| dc.subject | Affinity Labels | en |
| dc.subject | Alkaline Phosphatase/metabolism | en |
| dc.subject | Amino Acid Sequence | en |
| dc.subject | Apyrase/antagonists & inhibitors/*isolation & purification/metabolism | en |
| dc.subject | Chromatography, High Pressure Liquid | en |
| dc.subject | Chromatography, Ion Exchange | en |
| dc.subject | Electrophoresis, Polyacrylamide Gel | en |
| dc.subject | Humans | en |
| dc.subject | Hydrogen-Ion Concentration | en |
| dc.subject | Molecular Sequence Data | en |
| dc.subject | Placenta/*enzymology | en |
| dc.subject | Solubility | en |
| dc.subject | Species Specificity | en |
| dc.subject | Staining and Labeling | en |
| dc.title | Purification and properties of human placental ATP diphosphohydrolase | en |
| heal.abstract | ATP diphosphohydrolase activity (ATP-DPH) has been previously identified in the particulate fraction of human term placenta [Papamarcaki, T. & Tsolas, O. (1990) Mol. Cell. Biochem. 97, 1-8]. In the present study we have purified to homogeneity and characterized this activity. A 260-fold purification has been obtained by solubilization of the particulate fraction and subsequent chromatography on DEAE Sepharose CL-6B and 5'-AMP Sepharose 4B. The preparation has been shown to be free of alkaline phosphatase even though the placental extract is rich in this activity. The purified enzyme is a glycoprotein and migrates as a single broad band of 82 kDa on SDS/PAGE. The same band is obtained after photoaffinity labeling of the enzyme with 8-azido-[alpha-32P]ATP. The enzyme has a broad substrate specificity, hydrolyzing triphosphonucleosides and diphosphonucleosides but not monophosphonucleosides or other phosphate esters. The activity is dependent on the addition of divalent cations Ca2+ or Mg2+. The Km values for ATP and ADP were determined to be 10 microM and 20 microM, respectively. Maximum activity was found at pH 7.0-7.5 with ATP as substrate, and pH 7.5-8.0 with ADP. The enzymic activity is inhibited by NaN3, NaF, adenosine 5'-[beta,gamma-imido]triphosphate and adenosine 5'-[alpha,beta-methylene]triphosphate. Protein sequence analysis showed ATP-DPH to be N-terminally blocked. Partial internal amino acid sequence information was obtained after chymotryptic cleavage and identified a unique sequence with no significant similarity to known proteins. ATP-DPH activity has been reported to be implicated in the prevention of platelet aggregation, hydrolysing ADP to AMP and thus preventing blood clotting. | en |
| heal.access | campus | - |
| heal.fullTextAvailability | TRUE | - |
| heal.identifier.secondary | http://www.ncbi.nlm.nih.gov/pubmed/8529670 | - |
| heal.identifier.secondary | http://onlinelibrary.wiley.com/store/10.1111/j.1432-1033.1995.066_c.x/asset/j.1432-1033.1995.066_c.x.pdf?v=1&t=h03iyepe&s=6ce3709e6a83815e9ef4b7092207df23468e14d9 | - |
| heal.journalName | Eur J Biochem | en |
| heal.journalType | peer-reviewed | - |
| heal.language | en | - |
| heal.publicationDate | 1995 | - |
| heal.recordProvider | Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής | el |
| heal.type | journalArticle | - |
| heal.type.el | Άρθρο Περιοδικού | el |
| heal.type.en | Journal article | en |
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