Medical College of Wisconsin
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ADP-ribosylation of membrane proteins by bacterial toxins in the presence of NAD glycohydrolase. Biochim Biophys Acta 1988 Apr 28;954(1):65-72 PMID: 2833927

Pubmed ID

2833927

Abstract

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 J

Author

Jenifer Coburn PhD Professor in the Medicine department at Medical College of Wisconsin




Scopus

2-s2.0-0024298736   13 Citations

MESH terms used to index this publication - Major topics in bold

Adenosine Diphosphate Ribose
Animals
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
jenkins-FCD Prod-321 98992d628744e349846c2f62ac68f241d7e1ea70