Inhibition of NAD(+) glycohydrolase and ADP-ribosyl cyclase activities of leukocyte cell surface antigen CD38 by gangliosides

We have recently reported that gangliosides act as inhibitors of ADP-ribosyltransferases and NAD(+) glycohydrolases (NADase) of pertussis toxin and the C3 exoenzyme from Clostridium botulinum (Hara-Yokoyama, M., Hirabayashi, Y., Irie, F., Syuto, B., Moriishi, H., Susya, H., and Furuyama, S. (1995) J. Biol. Chem. 270, 8115-8121). Here, we investigated the effect of gangliosides on the enzymatic activity of leukocyte cell surface antigen CD38, which is identified as an ecto-NADase (Kontani, K., Nishina, H., Ohoka, Y., Takahashi, K., and Katada, T. (1993) J. Biol. Chem. 268, 16895-16898). Gangliosides G(M1a) and G(Q1b alpha) inhibited the NADase activity in the immunoprecipitate of anti-CD38 antibody from the membrane extract of retinoic acid-treated human leukemic HL-60 cells. Gangliosides also inhibited the NADase activity of the extracellular domain of CD38 antigen that was deprived of the transmembrane domain and was expressed in Escherichia coli as a fusion protein with maltose-binding protein (MBP-CD38). The order of the inhibitory effect of purified ganglioside species on the NADase activity on MBP-CD38 was as follows: G(Q1b alpha) > G(T1b), G(Q1b) > G(D1a), G(D1b), G(M1a),G(M1b), G(D3), G(M3). G(Q1b alpha) inhibited the NADase of MBP-CD38 in a noncompetitive manner versus NAD(+) with a K-i value of about 0.3 mu M. Neither ceramide nor the oligosaccharide moiety of G(Q1b alpha) had an effect on the NADase activity. G(Q1b alpha) G(T1b), and G(Q1b) also efficiently inhibited the ADP-ribosyl cyclase activity of MBP-CD38. At present, gangliosides are the only endogenous species that can block the enzymatic activity of CD38 antigen. The present results suggest a potential role of gangliosides as inhibitors of the ecto-NADases.