Requirements for antiribosomal activity of pokeweed antiviral protein.
It has been known for some time that pokeweed antiviral protein acts by enzymatically inhibiting protein synthesis on eucaryotic ribosome systems. The site of this action is known to be the ribosome itself. In this paper we show that the pokeweed antiviral protein reaction against ribosomes is a strong function of salt concentrations, where 160 mM K+ and 3 mM Mg2+ retards the reaction, while 20 mM K+ and 2 mM Mg2+ allows maximum reaction rate. It is also shown, however, that an unidentified protein in the postribosomal supernatant solution, together with ATP, allows the ribosome to be attacked even in the presence of high salt. Kinetic analysis of the antiviral protein reaction has been carried out under both sets of conditions, and reveals that the turnover number for the enzyme is about 300-400 mol/mol per min. in each case. The Km for ribosomes is 1 microM in the presence of low salt and 0.2 microM at higher salt in the presence of postribosomal supernatant factors plus ATP. The antiviral protein reaction is also shown to be pH dependent and is controlled by a residue with pKa value of approx. 7.0, apparently a histidine. Stoichiometric reaction of the enzyme with iodoacetamide results in a significant loss of antiribosomal activity.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009699:N-Glycosyl Hydrolases; D010940:Plant Proteins / Q000494:pharmacology*; D011188:Potassium / Q000494:pharmacology; D011817:Rabbits; D054789:Ribosome Inactivating Proteins, Type 1; D012270:Ribosomes / Q000187:drug effects*; D013997:Time Factors
D000998:Antiviral Agents; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D009699:N-Glycosyl Hydrolases; C024275:pokeweed antiviral protein; D008274:Magnesium; D011188:Potassium