Document Detail

Photoaffinity labeling of diphtheria toxin fragment A with NAD: structure of the photoproduct at position 148.
MedLine Citation:
PMID:  3864158     Owner:  NLM     Status:  MEDLINE    
Irradiation of mixtures of diphtheria toxin fragment A and [carbonyl-14C]NAD with UV light (253.7 nm) is known to induce efficient transfer of the radiolabel to position 148, corresponding to glutamic acid in the unmodified protein. Here we report the structure of the photoproduct at position 148, as determined by chemical and photochemical methods, fast-atom-bombardment mass spectrometry, and nuclear magnetic resonance. The photoproduct [an alpha-amino-gamma-(6-nicotin-amidyl)butyric acid residue] contains the entire nicotinamide moiety of NAD linked via its number 6 carbon to the decarboxylated gamma-methylene carbon of Glu-148. No portion of the ADP-ribosyl group of NAD is present. These findings are consistent with the idea that Glu-148 lies at or near the catalytic center of diphtheria toxin.
S F Carroll; J A McCloskey; P F Crain; N J Oppenheimer; T M Marschner; R J Collier
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  82     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1985 Nov 
Date Detail:
Created Date:  1985-12-05     Completed Date:  1985-12-05     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  7237-41     Citation Subset:  IM    
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MeSH Terms
ADP Ribose Transferases*
Affinity Labels
Bacterial Toxins*
Binding Sites
Diphtheria Toxin / analysis*,  radiation effects
Exotoxins / analysis,  radiation effects
Magnetic Resonance Spectroscopy
Mass Spectrometry
NAD / analysis*
Peptide Fragments / analysis*,  radiation effects
Protein Conformation
Ultraviolet Rays
Virulence Factors*
Grant Support
Reg. No./Substance:
0/Affinity Labels; 0/Bacterial Toxins; 0/Diphtheria Toxin; 0/Exotoxins; 0/Peptide Fragments; 0/Virulence Factors; 0/diphtheria toxin fragment A; 53-84-9/NAD; EC 2.4.2.-/ADP Ribose Transferases; EC protein, Pseudomonas aeruginosa

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