ADP-ribosylation of membrane proteins by bacterial toxins in the presence of NAD glycohydrolase. Biochim Biophys Acta 1988 Apr 28;954(1):65-72
Date
04/28/1988Pubmed ID
2833927DOI
10.1016/0167-4838(88)90056-8Scopus ID
2-s2.0-0024298736 (requires institutional sign-in at Scopus site) 15 CitationsAbstract
The ADP-ribosylation of membrane G proteins is difficult to achieve in tissues that are rich in membrane-bound NAD glycohydrolase (NAD+ glycohydrolase, EC 3.2.2.5). For many animal species this problem can be surmounted by inhibiting NAD hydrolysis with a combination of the anti-tuberculous drug, isonicotinic acid hydrazide, and the NAD analog, 3-acetylpyridine adenine dinucleotide, which act synergistically. In their presence, the ADP-ribosylation of cholera and pertussis toxin substrates reach plateau levels even with only 10 microM NAD. Although 3-acetylpyridine adenine dinucleotide acts as a weak substrate for the toxins, it is simple to estimate its contribution to the ADP-ribosylation and thus to determine the total amount of ADP-ribosylation substrate present in a tissue sample. NAD glycohydrolases that are insensitive to isonicotinic acid hydrazide are also less sensitive to 3-acetylpyridine adenine dinucleotide, but may be inactivated by dithiothreitol. Isonicotinic acid hydrazide adenine dinucleotide, the product of an exchange reaction catalysed by NAD glycohydrolase, runs with NAD in most thin-layer chromatographic systems. It can be separated from NAD, and quantitated, if the chromatographic solvent contains benzaldehyde. Isonicotinic acid hydrazide itself inhibits NAD glycohydrolase. It need not first be converted into isonicotinic acid hydrazide adenine dinucleotide.
Author List
Gill DM, Coburn JAuthor
Jenifer Coburn PhD Professor in the Medicine department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Adenosine Diphosphate RiboseAnimals
Brain Chemistry
Cattle
Cholera Toxin
Columbidae
Drug Synergism
Erythrocyte Membrane
GTP-Binding Proteins
Isoniazid
Membrane Proteins
Mice
NAD
NAD+ Nucleosidase
Pertussis Toxin
Swine
Virulence Factors, Bordetella
