Abstract
ADP-ribosylation, involving the transfer of an ADP-ribose moiety from NAD to proteins, is mediated by several bacterial toxins and endogenous ADP-ribosyltransferases. We report here the synthesis of biotinylated NAD and its use to label and purify biotinyl-ADP-ribosylated proteins. We demonstrate that biotinylated NAD can be used by diphtheria toxin to biotinylate elongation factor 2. Using avidin affinity chromatography, we have purified a protein whose ADP-ribosylation is enhanced by nitric oxide and which has been identified as glyceraldehyde-3-phosphate dehydrogenase.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Diphosphate Ribose / chemistry*
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Animals
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Biotin / chemistry*
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Brain Chemistry
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Chromatography, High Pressure Liquid
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Electrophoresis, Polyacrylamide Gel
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NAD / chemical synthesis*
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Nerve Tissue Proteins / chemistry
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Nerve Tissue Proteins / isolation & purification*
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Nitric Oxide / chemistry*
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Peptide Elongation Factor 2
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Peptide Elongation Factors / chemistry
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Peptide Elongation Factors / isolation & purification*
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Rats
Substances
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Nerve Tissue Proteins
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Peptide Elongation Factor 2
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Peptide Elongation Factors
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NAD
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Adenosine Diphosphate Ribose
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Nitric Oxide
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Biotin