Human placental (Asymmetrical) diadenosine 5',5'''-P1,P4-tetraphosphate hydrolase: purification to homogeneity and some properties.
The diadenosine 5',5'''-P1,P4-tetraphosphate (asymmetrical) hydrolase (EC 3.6.1.17) from human placenta has been purified to homogeneity by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephacel, gel filtration on Sephadex G-100, and affinity elution from red Sepharose. The enzyme is a single polypeptide of M(r) 19,200. It exhibits maximum (100%) activity at pH 7.3 in the presence of 3 mM MgCl2 and 60, 50, and 40% of the activity in 1 mM CoCl2, 0.1 mM ZnCl2, and 0.5 mM MnCl2, respectively. The Km value calculated for diadenosine tetraphosphate in the presence of Mg2+ is 10 microM and in the presence of Zn2+ 40 microM. Adenosine 5'-tetraphosphate, guanosine 5'-tetraphosphate, and fluoride proved to be inhibitors of the diadenosine tetraphosphate hydrolase; the I50 values were 6, 10, and 20 microM, respectively. Diguanosine tetraphosphate, bis-2,6-diaminopurine beta-D-ribofuranoside tetraphosphate, and diadenosine pentaphosphate were substrates for the hydrolase; relative velocities of hydrolysis estimated for 0.5 mM diadenosine tetraphosphate and these other substrates were 1:0.51:0.44:0.20, respectively. Diadenosine tetraphosphate analogues with P2-P3 bridges such as -CF2-, -CCl2-, and -CH2- were hydrolyzed to adenosine 5'-phosphate and the corresponding adenosine 5'-triphosphate analogue.(ABSTRACT TRUNCATED AT 250 WORDS)
Lazewska
D
D
Katedra Biochemii i Biotechnologii, Akademia Rolnicza, Poznań, Poland.
Starzyńska
E
E
Guranowski
A
A
D017766:Acid Anhydride Hydrolases; D002384:Catalysis; D002413:Cations, Divalent / Q000737:chemistry; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000737:chemistry; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D010920:Placenta / Q000201:enzymology*; D013379:Substrate Specificity
D002413:Cations, Divalent; D008670:Metals; D010744:Phosphoric Monoester Hydrolases; D017766:Acid Anhydride Hydrolases; C020023:bis(5'-nucleosyl)tetraphosphatase (asymmetrical)