Biomaterial Database

Ca2+, Mg2+-dependent endonuclease and ADP-ribosylation. 1983

K Yoshihara, and Y Tanaka, and T Kamiya

The molecular mechanism of the inhibition of Ca2+, Mg2+-dependent endonuclease by ADP-ribosylation was studied by using purified bull seminal plasma Ca2+, Mg2+-dependent endonuclease, endonuclease-stimulating proteins, and poly-(ADP-ribose) polymerase. The activity of an essentially homogeneous preparation of the endonuclease was markedly suppressed by its preincubation with NAD+, poly-(ADP-ribose) polymerase, DNA, and Mg2+. These four components of the incubation mixture were all essential for the suppression of the activity. Analyses of the initial and the chased reaction product by Sephadex G-100 column chromatography and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis revealed that Ca2+, Mg2+-dependent endonuclease was ADP-ribosylated during the incubation and its activity was markedly inhibited by the elongation of the ADP-ribose polymer covalently attached to the endonuclease. When the suppressed enzymes were mildly treated with an alkaline pH of 10.0, the activity was restored almost to the level of the unmodified control sample. These facts indicate that the linkage between the enzyme and poly(ADP-ribose) is hydrolyzed at this pH, and that the liberated polymer itself does not appreciably affect the endonuclease activity. These results also suggest that an electric repulsion between negative charges on DNA and poly(ADP-ribose) attached to Ca2+, Mg2+-dependent endonuclease is the basis for the observed suppression of the enzyme by ADP-ribosylation. Though histone H2B and H1 are shown to be as good endonuclease-stimulators (1) as they are good acceptors of ADP-ribose in poly(ADP-ribose) polymerase reaction (2), ADP-ribosylation of these two proteins did not affect their endonuclease-stimulating ability appreciably, at least under the conditions used.

UI	MeSH Term	Description	Entries
D008274	Magnesium	A metallic element that has the atomic symbol Mg, atomic number 12, and atomic weight 24.31. It is important for the activity of many enzymes, especially those involved in OXIDATIVE PHOSPHORYLATION.
D008297	Male		Males
D009702	Nucleoside Diphosphate Sugars		Diphosphate Sugars, Nucleoside,Sugars, Nucleoside Diphosphate
D011065	Poly(ADP-ribose) Polymerases	Enzymes that catalyze the transfer of multiple ADP-RIBOSE groups from nicotinamide-adenine dinucleotide (NAD) onto protein targets, thus building up a linear or branched homopolymer of repeating ADP-ribose units i.e., POLY ADENOSINE DIPHOSPHATE RIBOSE.	ADP-Ribosyltransferase (Polymerizing),Poly ADP Ribose Polymerase,Poly(ADP-Ribose) Synthase,Poly(ADP-ribose) Polymerase,PARP Polymerase,Poly ADP Ribose Transferase,Poly ADP-Ribose Synthase,Poly(ADP-Ribose) Transferase,Poly(ADPR) Polymerase,Poly(ADPribose) Polymerase,Poly ADP Ribose Synthase,Polymerase, PARP,Synthase, Poly ADP-Ribose
D011485	Protein Binding	The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.	Plasma Protein Binding Capacity,Binding, Protein
D002118	Calcium	A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes.	Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002417	Cattle	Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.	Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D004720	Endonucleases	Enzymes that catalyze the hydrolysis of the internal bonds and thereby the formation of polynucleotides or oligonucleotides from ribo- or deoxyribonucleotide chains. EC 3.1.-.	Endonuclease
D006657	Histones	Small chromosomal proteins (approx 12-20 kD) possessing an open, unfolded structure and attached to the DNA in cell nuclei by ionic linkages. Classification into the various types (designated histone I, histone II, etc.) is based on the relative amounts of arginine and lysine in each.	Histone,Histone H1,Histone H1(s),Histone H2a,Histone H2b,Histone H3,Histone H3.3,Histone H4,Histone H5,Histone H7
D006863	Hydrogen-Ion Concentration	The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH	pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations