Serine base exchange enzyme in porcine lyophilised platelets: enzyme properties and modulation by AlF4- and different types of heparin.
Phosphatidylserine is one of the PKC modulators and thus it may play an important role in signal transduction. Regulation of the synthesis of this phospholipid is not yet clarified. The contrasting reports are possibly related to the existence of different enzymes which, in mammalian tissues, catalyse the exchange between free serine and the nitrogen base of a membrane phospholipid. This study demonstrates that serine base exchange reactions of commercially available lyophilised porcine platelets exhibit similar pH optima, temperature and Ca2+ dependence as observed in fresh tissues. Analysis of fatty acids composition of the three phospholipid classes involved in base exchange reactions also demonstrated a similarity with fresh platelets. Serine and ethanolamine base exchange enzyme activities were assayed in parallel in platelet lysate subjected to preincubation at various temperatures (30-60 degrees C). When dithioerithrol was omitted from the incubation medium, the two base exchange reactions were inhibited with a similar temperature-dependent pattern. Addition of the reducing agent enhanced the sensitivity to preincubation only for the serine base exchange reaction which was inhibited by 80% after preincubation at 45 degrees C. With respect to its regulation, porcine platelet serine base exchange enzyme(s) was inhibited by fluoroalluminate, a widely used G-protein activator, and stimulated by unfractionated heparin. Low mol. wt. heparin did not influence enzyme activity. Unfractionated heparin greatly stimulated SBEE activity assayed at pH 7.4, a pH value far from the optimal pH.
Buratta
S
S
Istituto di Biochimica e Chimica Medica, Universita' di Perugia, Italy.
Andreoli
V
V
Mambrini
R
R
Iorio
A
A
Porcellati
S
S
Mozzi
R
R
D017607:Aluminum Compounds / Q000494:pharmacology*; D000818:Animals; D001792:Blood Platelets / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D005459:Fluorides / Q000494:pharmacology*; D005612:Freeze Drying; D006493:Heparin / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D019884:Nitrogenous Group Transferases / Q000378:metabolism*; D010718:Phosphatidylserines / Q000096:biosynthesis; D013552:Swine
D017607:Aluminum Compounds; D010718:Phosphatidylserines; D006493:Heparin; D019884:Nitrogenous Group Transferases; C022370:phospholipid serine base exchange enzyme; D005459:Fluorides; C032311:aluminum fluoride