title: 2'-Phosphoadenylylation of eukaryotic proteins: a type of covalent modification.
abstract: An enzymatic system in rat liver microsomal preparations has been detected that catalyzes the transfer of the 2'-phospho-AMP moiety from NADP to endogenous polypeptides; the major acceptor is a polypeptide of about 40 kDa (p40). Modification of the acceptor by 2'-phospho-AMP residues was deduced from the simultaneous transfer of 2'-[33P]phosphate and [3H]adenine residues from double-labeled NADP, while no incorporation of radioactivity into p40 was seen with NADP species labeled in the NMN moiety. The true substrate of this phosphoadenylylation reaction was 2'-phospho-ADP-ribose rather than NADP, because labeled phospho-ADP-ribose was as efficient as or more efficient than NADP in forming modified p40. Also, NADP was rapidly converted to phospho-ADP-ribose during incubation with microsomes. Furthermore, isonicotinic acid hydrazide, an inhibitor of NADP glycohydrolase, prevented phosphoadenylylation from NADP, but not from phospho-ADP-ribose, and glycohydrolase-resistant NADPH could not substitute for NADP. Transferase activity was found in liver and brain microsomes and, to a smaller extent, in the cytosol fractions. In Ehrlich ascites tumor cells, most of the activity resided in the cytosol, from which it could be partially purified. The apparent Km for phospho-ADP-ribose was about 2 X 10(-4) M, and the pH optimum was around 7. Divalent cations like Mg2+ and Mn2+ inhibited the reaction. In all compartmental preparations, activity was eliminated by heating or short treatment with alkali or acid. In submitochondrial particles from rat liver, a system with different characteristics led to the phosphoadenylylation of several endogenous polypeptides.
authors.forename: H
authors.initials: H
authors.lastname: Hilz
authors.forename: W
authors.initials: W
authors.lastname: Fanick
authors.forename: K
authors.initials: K
authors.lastname: Klapproth
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000031:analogs & derivatives* / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D002451:Cell Compartmentation; D055598:Chemical Phenomena; D002621:Chemistry; D008862:Microsomes, Liver / Q000378:metabolism*; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism*; D009702:Nucleoside Diphosphate Sugars; D010750:Phosphoproteins / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats
chemical_list: D009702:Nucleoside Diphosphate Sugars; D010750:Phosphoproteins; D000246:Adenosine Diphosphate Ribose; D000249:Adenosine Monophosphate; D009249:NADP