Arginine-Specific ADP-Ribosyltransferase and Its Acceptor Protein p33 in Chicken Polymorphonuclear Cells: Co-Localization in the Cell Granules, Partial Characterization, and In Situ Mono(ADP-Ribosyl)ation1

Publisher Website
Google Scholar
Abstract
We have reported the purification and characterization of arginine-specific ADP-ribo-syltransferase from hen liver nuclei [Tanigawa, Y. et al. (1984) J. BioL Chem. 259, 2022–2029] and the DNA-dependent mono(ADP-ribosyl)ation of p33, an acceptor protein in the nuclei [Mishima, K. et al. (1989) Eur. J. Biochem. 179, 267–273]. In the present study, we obtained evidence that among various tissues and cells from chicken, polymorphonuclear cells, so-called heterophils, possess both the ADP-ribosyltransferase and p33 at high levels. Percoll density gradient centrifugation of the postnuclear fraction of the heterophils revealed the co-localization of ADP-ribosyltransferase with p33 in the granule fraction. The enzyme and p33 were purified approximately 219- and 3.77-fold, respectively, from postnuclear pellet fraction to apparent homogeneity. The properties of heterophil ADP-ribosyltransferase and p33 were compared with those of the liver enzyme and p33. The molecular mass of the heterophil enzyme was estimated by SDS-polyacrylamide gel electrophoresis to be 27.5 kDa. The enzyme activity was stimulated by a sulfhydryl agent and inhibited by lysolecithin, NaCl, and inorganic phosphate. The mono(ADP-ribosyl)-ation of p33 was markedly enhanced by polyanion, such as DNA, RNA, or poly(L-gluta-mate). SDS-polyacrylamide gel electrophoretic analysis after limited trypsin proteolysis of p33s, purified from chicken heterophils and liver, showed much the same pattern. Thus, it appears that ADP-ribosyltransferase and p33 present in heterophils are identical to those in the liver, respectively. p33 is considered to be an in situ substrate for ADP-ribosyltransferase, since it was specifically mono(ADP-ribosyl)ated in permeabilized heterophils. Chicken ADP-ribosyltransferase may be involved in the function of polymorphonuclear cells through modification of the granule protein p33.

This publication has 0 references indexed in Scilit: