[Interaction of oligonucleotides and ATP with preparations of sIgA possessing protein kinase activity].
Interaction of secretory immunoglobulins A of a varying degree of purity with oligonucleotides and ATP has been studied by the method of affinity modification. For this aim we used reactive derivatives of 32P-labeled deoxyoligonucleotide (ClRCH2NHp(T)14) and [gamma-32P]ATP (ClR-32PppA) or ATP (ClR-pppA) bearing a 4-[(N-2-chloroethyl-N-methyl)amino]benzylamine residue. Preparations of sIgA were obtained from human milk by sequential chromatography on protein A-sepharose (P1), DEAE-fractogel (P2) and by gel-filtration in 50 mM NaOH (P3). It was revealed, that the H- and L-chains of sIgA P1; H-, L-chains and secretory component (SC) in sIgA P2 and only SC in sIgA P3 were exposed to modification after incubation with ClRCH2NHp(T)14. LPS, DNA, tRNA, heparin, sufficiently inhibited the modification of chains of sIgA P1. These competitors did not influence the modification of H- and L-chains of sIgA P2, but DNA, tRNA, heparin, inhibited binding of SC with the modifier. Suppressing affect of binding of ClRCH2NHp(T)14 with secretory component of sIgA P3 by d(T)14 has been observed as well. The research of ClR-32PppA interaction with sIgA P3 has shown that H- and L-chains of sIgA are exposed to modification. ATP inhibited the reaction. Study of the influence of modification on the protein kinase activity of sIgA P3 has revealed, that the preliminary incubation of sIgA P3 with ClR-pppA leads to inhibition of protein kinase activity. We suggest that sIgA, possessing the protein kinase activity (sIgA-abzymes) has an ATP-binding center (catalytic center) and has an oligonucleotide-binding center as well.
Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv. yuyakit@yahoo.com
Iu Ia
IuIa
Kit
L A
LA
Iakubov
E V
EV
Kuligina
V A
VA
Rikhter
D000255:Adenosine Triphosphate / Q000378:metabolism*; D002846:Chromatography, Affinity; D006801:Humans; D007071:Immunoglobulin A, Secretory / Q000302:isolation & purification / Q000378:metabolism*; D008895:Milk, Human / Q000201:enzymology*; D009841:Oligonucleotides / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*
D007071:Immunoglobulin A, Secretory; D009841:Oligonucleotides; D000255:Adenosine Triphosphate; D011494:Protein Kinases