The relationship between the binding of ATP and calcium to annexin IV. Effect of nucleotide on the calcium-dependent interaction of annexin with phosphatidylserine.
With the use of ATP analogues, we have found that porcine liver annexin (Anx) IV can be covalently labelled with 8-azido[gamma-32P]-ATP in the presence of Ca2+ (Kd 4.2 microM) and that the labelling is prevented by asolectin/cholesterol liposomes or chelation of calcium ions. On the other hand, non-covalent binding of 2'-(or 3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP) to AnxIV occurs optimally in the presence of liposomes and Ca2+ (Kd 7 microM). These observations were further confirmed by the results of intrinsic fluorescence quenching of AnxIV with various nucleotides, suggesting the existence of a relationship between Ca(2+)-, phospholipid- and ATP-binding sites within the annexin molecule. The interaction of AnxIV with nucleotides does not significantly affect its in vitro properties concerning the binding to phosphatidylserine (PS) monolayers.
Bandorowicz-Pikula
J
J
Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland.
Wrzosek
A
A
Makowski
P
P
Pikula
S
S
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D000345:Affinity Labels / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D017314:Annexin A4 / Q000378:metabolism*; D001386:Azides / Q000378:metabolism / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D009705:Nucleosides / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D013329:Structure-Activity Relationship; D013552:Swine
D000345:Affinity Labels; D017314:Annexin A4; D001386:Azides; D005456:Fluorescent Dyes; D009705:Nucleosides; D010718:Phosphatidylserines; D010761:Phosphorus Radioisotopes; C017040:8-azidoadenosine 5'-triphosphate; C045674:2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D002118:Calcium