Purification and characterization of ADP-ribosyl cyclase from Euglena gracilis.
ADP-ribosyl cyclase, which catalyzes the conversion from NAD+ to cyclic adenosine diphosphoribose (cADPR), is proposed to participate in cell cycle regulation in Euglena gracilis. This enzyme, which was found as a membrane-bound protein, was purified almost the homogeneity after solubilization with deoxycholate, and found to be a monomeric protein with a molecular mass of 40 kDa. Its Km value for NAD+ was estimated to be 0.4 mM, and cADPR, a product of the enzyme, inhibited the enzyme competitively with respect to NAD+ whereas another product, nicotinamide, showed noncompetitive (mixed-type) inhibition. In contrast to mammalian CD38 and BST-1, Euglena ADP-ribosyl cyclase lacked cADPR hydrolase activity.
Masuda
W
W
Department of Applied Biological Chemistry, Osaka Prefecture University, Sakai, Osaka, 599-8531, Japan.
Takenaka
S
S
Tsuyama
S
S
Inui
H
H
Miyatake
K
K
Nakano
Y
Y
D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000302:isolation & purification* / Q000378:metabolism*; D002453:Cell Cycle; D005056:Euglena gracilis / Q000166:cytology / Q000201:enzymology*; D009244:NAD+ Nucleosidase / Q000302:isolation & purification* / Q000378:metabolism*
D015703:Antigens, CD; D000943:Antigens, Differentiation; D036541:ADP-ribosyl Cyclase; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1