Nitric Oxide-Induced Modification of Glyceraldehyde-3-Phosphate Dehydrogenase with NAD+ Is Not ADP-Ribosylation

Abstract

One biological effect of nitric oxide (NO) has been believed to be exerted through induction of the ADP-ribosyltransferase activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Though this notion is based on the finding that NO increases the auto-ADP-ribosylation of GAPDH, controversial data have also been reported. To determine whether or not NO really activates ADP-ribosylation, we re-examined the NO-induced modification of GAPDH with NAD+. GAPDH was modified equally with [adenosine-14C]NAD+ and [carbonyl-14C]NAD+, indicating that the glycoside bond of NAD+ between ADP-ribose and nicotinamide is intact. The release of nicotinamide from NAD+ was not evident during incubation of GAPDH with [carbonyl-14C]NAD+. Thus, the modification of GAPDH is apparently not ADP-ribosylation. In addition, we found that basal and glyceraldehyde-3-phosphate-induced modifications of GAPDH, both of which have also been explained as ADP-ribosylation, were not ADP-ribosylation, and that the modification of GAPDH in the absence and presence of NO or GA3P was distinct in the dithiothreitol effect or resistance to HgCl2.

Mouse Sir2 Homolog SIRT6 Is a Nuclear ADP-ribosyltransferase

“…Such labeling, as observed in the case of glyceraldehyde-3-phosphate dehydrogenase (29), can be misinterpreted as ADP-ribosylation of specific amino acid acceptors (29,30). We have observed low levels of nonspecific radiolabeling of proteins incubated in the presence of [ 32 P]NAD ϩ .…”

Glyceraldehyde-3-phosphate dehydrogenase is negatively regulated by ADP-ribosylation in the fungus Phycomyces blakesleeanus

“…These authors also reported that NOmediated ADP-ribosylation of GAPDH inhibited this enzyme activity. Several investigators have questioned whether or not auto-ADP-ribosylation of GAPDH occurs as a result of stimulation by NO (McDonald & Moss, 1993 ;Itoga et al, 1997).…”

Intracellular Redox State Determines Whether Nitric Oxide Is Toxic or Protective to Rat Oligodendrocytes in Culture

“…In the last several years, several studies have been published, showing a protective effect of nitric oxide in a variety of paradigms of cell injury and cell death. These include (1) direct scavenging of free radicals (Hogg et al, 1993 ; Wink et al, 1993) ; (2) effects on intracellular iron metabolism, including interaction with iron to form nitrosyl‐iron complexes (Rauhala et al., 1996 ; Sergent et al, 1997 ; Yoshie and Ohshima, 1997), preventing release of iron from ferritin (Puntarulo and Cederbaum, 1997), or stimulating iron‐responsive RNA binding elements such as cis ‐aconitase (Jaffrey et al, 1994) ; (3) inactivation of caspases (Dimmeler et al, 1997) ; (4) activation of a cyclic GMP‐dependent survival pathway, as has been seen in PC12 cells (Farinelli et al, 1996) and tumor necrosis factor‐α‐induced apoptosis in endothelial cells (Polte et al, 1997) ; (5) inducing expression of protective proteins such as heat shock proteins (Kim et al, 1997) ; (6) inhibition of nuclear factor‐κB activation (Peng et al, 1995 ; Matthews et al, 1996 ; Park et al, 1997 ; Togashi et al, 1997) ; (7) inhibition of glyceradehyde‐3‐phosphate dehydrogenase (Dimmeler et al, 1992 ; Kots et al, 1992 ; Molina y Vedia et al, 1992 ; McDonald and Moss, 1993 ; Itoga et al, 1997), whose activity appears to be required in one paradigm of neuronal apoptosis (Ishitani et al, 1996) ; and (8) oxidation in neurons of a redox modulatory site on the NMDA receptor, resulting in a decrease in NMDA receptor‐mediated currents (Lei et al, 1992), a mechanism that remains controversial (Aizenman et al, 1998).…”