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Article: Caveat: Mycoplasma arginine deiminase masquerading as nitric oxide synthase in cell cultures

TitleCaveat: Mycoplasma arginine deiminase masquerading as nitric oxide synthase in cell cultures
Authors
KeywordsSpecies Index: Canis Familiaris
Mycoplasma
Primates
Issue Date1998
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/bbamcr
Citation
Biochimica Et Biophysica Acta - Molecular Cell Research, 1998, v. 1404 n. 3, p. 314-320 How to Cite?
AbstractWe used confluent cultures of dog gallbladder epithelial cells, stimulated by conditioned medium from a culture of human neonatal foreskin fibroblasts, to establish the presence of inducible nitric oxide synthase (NOS, EC 1.14.13.39). Assay was by conversion of radiolabeled arginine to citrulline. By 4 days after addition of the conditioned medium, a relatively high level of activity was observed. However, further study showed that the enzyme did not require addition of the usual cofactors for maximal activity (NADPH, FAD, FMN and tetrahydrobiopterin) and was stable in the absence of anti-proteolytic agents. Our suspicion that this enzyme might not be NOS but arginine deiminase (EC 3.5.3.6) was confirmed by enzyme purification and by the liberation of ammonia during enzyme reaction. This enzyme, which is absent from primates and virtually confined to single-cell organisms, suggested the presence of Mycoplasma, a common contaminant of cell cultures, and it was subsequently confirmed that the fibroblast culture was a source of Mycoplasma. With the widespread interest in nitric oxide and NOS, and common use of the convenient [3H]arginine assay, there is a considerable danger of the two enzymes being confused. At the very least, it is necessary to check for activity in the absence of added cofactors. Copyright (C) 1998 Elsevier Science B.V.
Persistent Identifierhttp://hdl.handle.net/10722/92516
ISSN
2015 Impact Factor: 5.128
2015 SCImago Journal Rankings: 3.043
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChoi, JWen_HK
dc.contributor.authorHaigh, WGen_HK
dc.contributor.authorLee, SPen_HK
dc.date.accessioned2010-09-17T10:48:37Z-
dc.date.available2010-09-17T10:48:37Z-
dc.date.issued1998en_HK
dc.identifier.citationBiochimica Et Biophysica Acta - Molecular Cell Research, 1998, v. 1404 n. 3, p. 314-320en_HK
dc.identifier.issn0167-4889en_HK
dc.identifier.urihttp://hdl.handle.net/10722/92516-
dc.description.abstractWe used confluent cultures of dog gallbladder epithelial cells, stimulated by conditioned medium from a culture of human neonatal foreskin fibroblasts, to establish the presence of inducible nitric oxide synthase (NOS, EC 1.14.13.39). Assay was by conversion of radiolabeled arginine to citrulline. By 4 days after addition of the conditioned medium, a relatively high level of activity was observed. However, further study showed that the enzyme did not require addition of the usual cofactors for maximal activity (NADPH, FAD, FMN and tetrahydrobiopterin) and was stable in the absence of anti-proteolytic agents. Our suspicion that this enzyme might not be NOS but arginine deiminase (EC 3.5.3.6) was confirmed by enzyme purification and by the liberation of ammonia during enzyme reaction. This enzyme, which is absent from primates and virtually confined to single-cell organisms, suggested the presence of Mycoplasma, a common contaminant of cell cultures, and it was subsequently confirmed that the fibroblast culture was a source of Mycoplasma. With the widespread interest in nitric oxide and NOS, and common use of the convenient [3H]arginine assay, there is a considerable danger of the two enzymes being confused. At the very least, it is necessary to check for activity in the absence of added cofactors. Copyright (C) 1998 Elsevier Science B.V.en_HK
dc.languageengen_HK
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/bbamcren_HK
dc.relation.ispartofBiochimica et Biophysica Acta - Molecular Cell Researchen_HK
dc.subjectSpecies Index: Canis Familiarisen_HK
dc.subjectMycoplasmaen_HK
dc.subjectPrimatesen_HK
dc.titleCaveat: Mycoplasma arginine deiminase masquerading as nitric oxide synthase in cell culturesen_HK
dc.typeArticleen_HK
dc.identifier.emailLee, SP: sumlee@hku.hken_HK
dc.identifier.authorityLee, SP=rp01351en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/S0167-4889(98)00079-2en_HK
dc.identifier.pmid9739159-
dc.identifier.scopuseid_2-s2.0-0032538007en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032538007&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume1404en_HK
dc.identifier.issue3en_HK
dc.identifier.spage314en_HK
dc.identifier.epage320en_HK
dc.identifier.isiWOS:000076189500005-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridChoi, JW=24477421700en_HK
dc.identifier.scopusauthoridHaigh, WG=6603814152en_HK
dc.identifier.scopusauthoridLee, SP=7601417497en_HK

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