Nucleotidase activity and Ca-uptake were characterized in endoplasmic reticulum (ER) enriched rat submandibular gland (SMG) microsomal preparations. (i) Ca-uptake had characteristics of an ER Ca-ATPase. (ii) Nucleotidase activity was equally stimulated by calcium, magnesium and manganese, but with different Km values. (iii) Specific inhibitors of P-type Ca-ATPases were ineffective on nucleotidase activity, demonstrating that this activity was not related to calcium uptake and did not correspond to classical Ca2+ pumps. (iv) ATP and UTP were more efficient substrates, whereas ADP and UDP were hydrolyzed at significantly slower rate. (v) Nucleotidase activity was sensitive to mild detergent solubilization and insensitive to ionophore addition. (vi) Nucleotidase activity was strongly inhibited by suramin, a nucleoside triphosphate diphosphohydrolase (NTPDase) inhibitor. (vii) Nucleotidase activity exponentially diminished as function of time. All these observations are consistent with a NTPDase identity. The presence of a NTPDase was demonstrated by immunohistochemistry in rat SMG. Immunoreactivity was stronger in ductal cells than in mucous and serous acini. Although this enzyme was observed in the plasma membrane, colocalization with the ER marker calnexin revealed a specific subcellular localization in this organelle of all three types of cell. The putative function of this NTPDase activity in salivary glands is discussed.
- MeSH
- adenosindifosfát metabolismus MeSH
- adenosintrifosfát metabolismus MeSH
- elektronová mikroskopie MeSH
- endoplazmatické retikulum enzymologie účinky léků MeSH
- financování organizované MeSH
- hořčík metabolismus MeSH
- hydrolýza MeSH
- imunohistochemie MeSH
- inhibitory enzymů farmakologie MeSH
- kinetika MeSH
- krysa rodu rattus MeSH
- mangan metabolismus MeSH
- mikrozomy enzymologie MeSH
- nukleotidasy antagonisté a inhibitory metabolismus MeSH
- podčelistní slinná žláza metabolismus účinky léků MeSH
- potkani Wistar MeSH
- sarkoplazmatická Ca2+-ATPáza metabolismus MeSH
- uridindifosfát metabolismus MeSH
- uridintrifosfát metabolismus MeSH
- vápník metabolismus MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
