{"refrec":{"BRefID":261811,"RR":"<b>Wilquet, V.; Gaspar, J.A.; Van de Lande, M.; Van de Casteele, M.; Legrain, C.; Meiering, E.M.; Glansdorff, N.</b> (1998). Purification and characterization of recombinant <i>Thermotoga maritima</i> dihydrofolate reductase. <i>Eur. J. Biochem. 255(3)</i>: 628-637. <a href=\"http://dx.doi.org/10.1046/j.1432-1327.1998.2550628.x\" target=\"_blank\">dx.doi.org/10.1046/j.1432-1327.1998.2550628.x</a>","BEntID":253829,"PublicFlag":1,"CheckedFlag":1,"wosflag":null,"vabbflag":null,"RefStringPartII":". <i>Eur. J. Biochem. 255(3)</i>: 628-637. <a href=\"https://dx.doi.org/10.1046/j.1432-1327.1998.2550628.x\" target=\"_blank\">https://dx.doi.org/10.1046/j.1432-1327.1998.2550628.x</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Wilquet, V.; Gaspar, J.A.; Van de Lande, M.; Van de Casteele, M.; Legrain, C.; Meiering, E.M.; Glansdorff, N.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Wilquet, V. <i>et al.</i>","Englishabstract":"We have overexpressed the gene for dihydrofolate reductase (DHFR) from <i>Thermotoga maritima</i> in <i>Escherichia coli</i> and characterized the biochemical properties of the recombinant protein. This enzyme is involved in the <i>de novo</i> synthesis of deoxythymidine 5′-phosphate and is critical for cell growth. High levels of <i>T. maritima</i> DHFR in the new expression system conferred resistance to high levels of DHFR inhibitors which inhibit the growth of non-recombinant cells. The enzyme was purified to homogeneity in the following two steps&nbsp;: heat treatment followed by affinity chromatography or cation-exchange chromatography. Most of the biochemical properties of <i>T. maritima</i> DHFR resemble those of other bacterial or eukaryotic DHFRs, however, some are unique to <i>T. maritima</i> DHFR. The pH optima for activity, <i>K</i><sub>m</sub> for substrates, and polypeptide chain length of <i>T. maritima</i> DHFR are similar to those of other DHFRs. In addition, the secondary structure of <i>T. maritima</i> DHFR, as measured by circular dichroism, is similar to that of other DHFRs. Interestingly, <i>T. maritima</i> DHFR exhibits some characteristics of eukaryotic DHFRs, such as a basic pI, an excess of positively charged residues in the polypeptide chain and activation of the enzyme by inorganic salts and urea. Unlike most other DHFRs which are monomeric or part of a bifunctional DHFR-thymidylate synthase (TS) enzyme, <i>T. maritima</i> DHFR seems to generally form a dimer in solution and is also much more thermostable than other DHFRs. It may be that dimer formation is a key factor in determining the stability of <i>T. maritima</i> DHFR.","AbstractOtherLang":null,"BibLvlCode":"AS","StandardTitle":"Purification and characterization of recombinant <i>Thermotoga maritima</i> dihydrofolate reductase","OrigTitleLangCode":"en","OrigTitleLangCodeExtended":"eng","OrigTitleLangID":15,"DateLastModified":{"date":"2024-12-10 01:33:17.368041","timezone_type":1,"timezone":"+01:00"},"UserAccessRight":null,"UserAccID":null,"AuthorKeywords":"dihydrofolate reductase; Thermotoga maritima; thermal stability;purification; expression in Escherichia coli","OtherDescriptors":null,"Notes":null,"AnaPub":1998,"MonPub":null,"DateUpdate":"2016-10-10","DateCreate":"2016-10-03","SecASFANote":null,"ConfID":null,"PeerRev":1,"VlizCoreFlag":1,"WoScode":"WOS:000075287900013","VABBcode":null,"OpenAcc":1,"DOI":"10.1046/j.1432-1327.1998.2550628.x"},"refs":null,"anarec":{"AnaID":261811,"PubliDate":1998,"Pagination":"628-637","XtraPublOfAnaID":null,"ISBN":null,"Volume":"255","Issue":"3","BRefMon":null,"BRefMonRR":null,"BRefXtra":null,"BRefXtraRR":null,"SerBRefID":42671,"SerRR":"European Journal of Biochemistry. Springer: Berlin.  ISSN 0014-2956; e-ISSN 1432-1033","StandardTitleSer":"European Journal of Biochemistry","ISSN":"0014-2956","AbbrevSer":"Eur. J. Biochem.","StandardTitleMon":null,"StartPage":628,"Pages":10,"ToPubliDate":null,"BRefBibLvlCode":"S","SerNotes":null},"monrec":null,"serrec":null,"relations":null,"relationsRev":null,"addrec":null,"othpubs":null,"ownerships":null,"authors":[{"AutName":"Wilquet","Firstname":"Valérie","Initials":"V.","Affiliation":null,"Discriminator":null,"CorporateFlag":0,"BEntID":253829,"AutID":242480,"OrderNr":1,"DegrID":null,"EditorFlag":0,"CorrespFlag":0,"IllustratorFlag":0,"ReviserFlag":0,"TranslatorFlag":0,"InsAcronym":null,"InsFSN":null,"ORCID":null,"PersID":34109,"InsID":null},{"AutName":"Gaspar","Firstname":"Joe","Initials":"J.A.","Affiliation":null,"Discriminator":null,"CorporateFlag":0,"BEntID":253829,"AutID":238899,"OrderNr":2,"DegrID":null,"EditorFlag":0,"CorrespFlag":0,"IllustratorFlag":0,"ReviserFlag":0,"TranslatorFlag":0,"InsAcronym":null,"InsFSN":null,"ORCID":null,"PersID":null,"InsID":null},{"AutName":"Van de Lande","Firstname":"Monica","Initials":"M.","Affiliation":null,"Discriminator":null,"CorporateFlag":0,"BEntID":253829,"AutID":238900,"OrderNr":3,"DegrID":null,"EditorFlag":0,"CorrespFlag":0,"IllustratorFlag":0,"ReviserFlag":0,"TranslatorFlag":0,"InsAcronym":null,"InsFSN":null,"ORCID":null,"PersID":null,"InsID":null},{"AutName":"Van de Casteele","Firstname":"Mark","Initials":"M.","Affiliation":null,"Discriminator":null,"CorporateFlag":0,"BEntID":253829,"AutID":242481,"OrderNr":4,"DegrID":null,"EditorFlag":0,"CorrespFlag":0,"IllustratorFlag":0,"ReviserFlag":0,"TranslatorFlag":0,"InsAcronym":null,"InsFSN":null,"ORCID":null,"PersID":null,"InsID":null},{"AutName":"Legrain","Firstname":"Christianne","Initials":"C.","Affiliation":null,"Discriminator":null,"CorporateFlag":0,"BEntID":253829,"AutID":242482,"OrderNr":5,"DegrID":null,"EditorFlag":0,"CorrespFlag":0,"IllustratorFlag":0,"ReviserFlag":0,"TranslatorFlag":0,"InsAcronym":null,"InsFSN":null,"ORCID":null,"PersID":null,"InsID":null},{"AutName":"Meiering","Firstname":"Elizabeth","Initials":"E.M.","Affiliation":null,"Discriminator":null,"CorporateFlag":0,"BEntID":253829,"AutID":238901,"OrderNr":6,"DegrID":null,"EditorFlag":0,"CorrespFlag":0,"IllustratorFlag":0,"ReviserFlag":0,"TranslatorFlag":0,"InsAcronym":null,"InsFSN":null,"ORCID":null,"PersID":null,"InsID":null},{"AutName":"Glansdorff","Firstname":"Nicolas","Initials":"N.","Affiliation":null,"Discriminator":null,"CorporateFlag":0,"BEntID":253829,"AutID":242483,"OrderNr":7,"DegrID":null,"EditorFlag":0,"CorrespFlag":0,"IllustratorFlag":0,"ReviserFlag":0,"TranslatorFlag":0,"InsAcronym":null,"InsFSN":null,"ORCID":null,"PersID":null,"InsID":null}],"mapdetails":null,"datasets":null,"monographs":null,"monparts":null,"serparts":null,"BEntOpen":null,"BEntPrivate":null,"availability":[{"BInstID":295833,"LibID":36,"BRefID":261811,"EmbargoDate":null,"FullEmbargoDate":null,"PhysMedID":16,"hasOCRd":1,"ShelfLocCode":"295833","RFID":null,"PaidValue":null,"Medium":"Server","Description":"VLIZ Open Access","Acronym":"VLIZ","Library":"Vlaams Instituut voor de Zee","DutchTerm":"Open access","URL":null,"ClassifID":53,"Classification":"Open access","ReqLink":null,"ClassifTypID":1,"URLLocation":"https://www.vliz.be/imisdocs/publications/","SubDir":null,"InternalReq":0,"LoggedInReq":0,"Disclaimer":null,"DutchDisclaimer":null,"FileFormat":".pdf","FileDescr":"pdf","InsPub":1,"InsID":36,"FileFormID":6,"LendableFlag":1,"PublicFlag":1,"orderLib":"A","Notes":null,"AccConID":null,"AccessConstraint":null,"LicURL":null}],"litstyles":null,"thespers":null,"arch2discl":null,"SERpubls":[{"PublName":"Springer","City":"Berlin"}],"MONpubls":null,"pictures":[],"thestermsPath":null,"thestermsASFA":null,"taxtermsASFA":[{"TaxTerm":"Escherichia coli"},{"TaxTerm":"Thermotoga maritima"}],"geotermsASFA":null,"collections":[{"Collection":"OMA - Open Marien Archief","ShortName":"OMA"}],"conf":null,"proj":null,"Physdatasets":null,"spcols":{"222":{"SpName":"BMB - Belgische Mariene Bibliografie","SpColID":222,"ParSpColID":null,"TopParID":null,"ShortName":"BMB","URLLocation":null,"LibID":36,"OpenRepoFlag":null,"SpTypID":null,"TopParIDNotWebsite":null,"SpColPath":"BMB"},"221":{"SpName":"OMA - Open Marien Archief","SpColID":221,"ParSpColID":null,"TopParID":null,"ShortName":"OMA","URLLocation":null,"LibID":36,"OpenRepoFlag":1,"SpTypID":null,"TopParIDNotWebsite":null,"SpColPath":"OMA"}},"doi":null,"publs":null,"serparttypes":null,"monauthors":null,"MParts":null,"SParts":null,"hLibs":null,"langs":[{"BEntID":253829,"AbstractFlag":0,"LangID":15,"LangCode":"en","Lang":"English","DutchTerm":"Engels","LangCodeExtended":"eng"},{"BEntID":253829,"AbstractFlag":1,"LangID":15,"LangCode":"en","Lang":"English","DutchTerm":"Engels","LangCodeExtended":"eng"}],"urls":[{"URL":"https://dx.doi.org/10.1046/j.1432-1327.1998.2550628.x","externalID":"10.1046/j.1432-1327.1998.2550628.x","URLTypeCode":"DOI","URLID":50520,"URLTypID":13,"URLType":"DOI","URLPrefix":"http://dx.doi.org/"}],"thesterms":null,"taxterms":[{"TaxTerm":"Escherichia coli","AphiaID":422817,"TaxtID":23081},{"TaxTerm":"Thermotoga maritima","AphiaID":567396,"TaxtID":42835}],"geoterms":null,"othterms":null,"asfacodes":null,"asfa2codes":null,"thestermsFRIS":null,"taxtermsFRIS":[{"TaxTerm":"Escherichia coli","DutchTerm":null,"AphiaID":422817,"TaxtID":23081},{"TaxTerm":"Thermotoga maritima","DutchTerm":null,"AphiaID":567396,"TaxtID":42835}],"geotermsFRIS":null,"othtermsFRIS":null,"resmessage":"","complete":1,"sessions":{"newSesName":"Lyssens, Liesbeth, L.","newSesDate":{"date":"2016-10-03 20:16:18.820000","timezone_type":3,"timezone":"Europe/Brussels"},"updSesName":"Bouchti, Zohra, Z.","updSesDate":{"date":"2016-10-10 09:21:13.483000","timezone_type":3,"timezone":"Europe/Brussels"}}}