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Article: NAD glycohydrolase and ADP-ribosyltransferase activities are intrinsic to the A 1 peptide of choleragen

TitleNAD glycohydrolase and ADP-ribosyltransferase activities are intrinsic to the A 1 peptide of choleragen
Authors
Issue Date1979
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal Of Biological Chemistry, 1979, v. 254 n. 23, p. 11993-11996 How to Cite?
AbstractIn prior reports (Moss, J., Manganiello, V. C., and Vaughan, M. (1976) Proc. Natl. Acd Sci. U. S. A. 73, 4424-4427), it was demonstrated that choleragen preparations possessed NAD glycohydrolase activity. Tait and Van Heyningen ((1978) Biochem. J. 174,1059-1062) have recently proposed that the activity observed in these preparations was due to a contaminating enzyme and was not an intrinsic activity of choleragen. The absence of an NAD glycohydrolase activity in choleragen preparation would imply that nicotinamide release from NM requires a ternary complex of NAD, acceptor protein, and toxin and that, in contrast to diphtheria toxin and Pseudomonas exotoxin A, choleragen cannot use water as an ADP-ribose acceptor. Since the ability to hydrolyze NAD is critical to understanding the mechanism of the toxin-catalyzed reaction, we have re-examined the question using purified choleragen peptides. In agreement with our prior observations, the purified A1 peptide of choleragen, but not the AZ or B, catalyzed the hydrolysis of NAD to ADP-ribose and nicotinamide and the NAD-dependent ADP-ribosylation of arginine. The NAD glycohydrolase activity cochromatographed with the A1 peptide, the ADP-ribosyltransferase activity, and the ability to activate rat liver adenylate cyclase. The K, for NAD was similar for the holotoxin and A1. The turnover number of A1 was similar to that of the intact toxin which contains one A1 per molecule. In contrast to the holotoxin or its A subunit, the A1 peptide did not exhibit a lag in reaction rate or a thiol requirement for activity. The enzymatic activity of the A1 peptide was inhibited by sodium dodecyl sulfate as was its ability to activate adenylate cyclase. These results support the conclusion that the A1 peptide of choleragen possesses NAD glycohydrolase activity and can activate the ribosyl-nicotinamide bond of NAD in the absence of an acceptor protein.
Persistent Identifierhttp://hdl.handle.net/10722/167445
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorMoss, Jen_US
dc.contributor.authorStanley, SJen_US
dc.contributor.authorLin, MCen_US
dc.date.accessioned2012-10-08T03:07:05Z-
dc.date.available2012-10-08T03:07:05Z-
dc.date.issued1979en_US
dc.identifier.citationJournal Of Biological Chemistry, 1979, v. 254 n. 23, p. 11993-11996en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/10722/167445-
dc.description.abstractIn prior reports (Moss, J., Manganiello, V. C., and Vaughan, M. (1976) Proc. Natl. Acd Sci. U. S. A. 73, 4424-4427), it was demonstrated that choleragen preparations possessed NAD glycohydrolase activity. Tait and Van Heyningen ((1978) Biochem. J. 174,1059-1062) have recently proposed that the activity observed in these preparations was due to a contaminating enzyme and was not an intrinsic activity of choleragen. The absence of an NAD glycohydrolase activity in choleragen preparation would imply that nicotinamide release from NM requires a ternary complex of NAD, acceptor protein, and toxin and that, in contrast to diphtheria toxin and Pseudomonas exotoxin A, choleragen cannot use water as an ADP-ribose acceptor. Since the ability to hydrolyze NAD is critical to understanding the mechanism of the toxin-catalyzed reaction, we have re-examined the question using purified choleragen peptides. In agreement with our prior observations, the purified A1 peptide of choleragen, but not the AZ or B, catalyzed the hydrolysis of NAD to ADP-ribose and nicotinamide and the NAD-dependent ADP-ribosylation of arginine. The NAD glycohydrolase activity cochromatographed with the A1 peptide, the ADP-ribosyltransferase activity, and the ability to activate rat liver adenylate cyclase. The K, for NAD was similar for the holotoxin and A1. The turnover number of A1 was similar to that of the intact toxin which contains one A1 per molecule. In contrast to the holotoxin or its A subunit, the A1 peptide did not exhibit a lag in reaction rate or a thiol requirement for activity. The enzymatic activity of the A1 peptide was inhibited by sodium dodecyl sulfate as was its ability to activate adenylate cyclase. These results support the conclusion that the A1 peptide of choleragen possesses NAD glycohydrolase activity and can activate the ribosyl-nicotinamide bond of NAD in the absence of an acceptor protein.-
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.subject.meshAdenosine Diphosphate Riboseen_US
dc.subject.meshAdenylate Cyclase - Metabolismen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCell Membrane - Drug Effects - Enzymologyen_US
dc.subject.meshCholera Toxin - Pharmacologyen_US
dc.subject.meshEnzyme Activationen_US
dc.subject.meshLiver - Enzymologyen_US
dc.subject.meshNaden_US
dc.subject.meshNad+ Nucleosidase - Metabolismen_US
dc.subject.meshNiacinamideen_US
dc.subject.meshNucleotidyltransferases - Metabolismen_US
dc.subject.meshRatsen_US
dc.titleNAD glycohydrolase and ADP-ribosyltransferase activities are intrinsic to the A 1 peptide of choleragenen_US
dc.typeArticleen_US
dc.identifier.emailLin, MC:mcllin@hkucc.hku.hken_US
dc.identifier.authorityLin, MC=rp00746en_US
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.pmid227885-
dc.identifier.scopuseid_2-s2.0-0018582776en_US
dc.identifier.volume254en_US
dc.identifier.issue23en_US
dc.identifier.spage11993en_US
dc.identifier.epage11996en_US
dc.identifier.isiWOS:A1979HX41200038-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridMoss, J=35373323000en_US
dc.identifier.scopusauthoridStanley, SJ=7202073665en_US
dc.identifier.scopusauthoridLin, MC=7404816359en_US

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