| pubmed-article:15249050 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:15249050 | lifeskim:mentions | umls-concept:C0314836 | lld:lifeskim |
| pubmed-article:15249050 | lifeskim:mentions | umls-concept:C0340305 | lld:lifeskim |
| pubmed-article:15249050 | lifeskim:mentions | umls-concept:C1998793 | lld:lifeskim |
| pubmed-article:15249050 | lifeskim:mentions | umls-concept:C1880022 | lld:lifeskim |
| pubmed-article:15249050 | pubmed:issue | 2 | lld:pubmed |
| pubmed-article:15249050 | pubmed:dateCreated | 2004-7-13 | lld:pubmed |
| pubmed-article:15249050 | pubmed:abstractText | The gene from Aeromonas veronii bv. sobria encoding the metallo-beta-lactamase ImiS was subcloned into pET-26b, and ImiS was over-expressed in BL21(DE3) Escherichia coli and purified using SP-Sepharose chromatography. This protocol yielded over 5 mg of ImiS per liter of growth culture under optimum conditions. The biochemical properties of recombinant ImiS were compared with those of native ImiS. Recombinant and native ImiS have the same N-terminus of A-G-M-S-L, and CD spectroscopy was used to show that the enzymes have similar secondary structures. Gel filtration chromatography revealed that both enzymes exist as monomers in solution. MALDI-TOF mass spectra showed that the enzymes have a molecular mass of 25,247 Da, and metal analyses demonstrated that both as-isolated enzymes bind ca. 0.7 mol of Zn(II). Metal titrations demonstrate that the maximum activity of recombinant ImiS occurs when the enzyme binds one equivalent of zinc. Steady-state kinetic studies reveal that recombinant ImiS is a carbapenemase like native ImiS and that the recombinant enzyme exhibits similar kcat and K(m) values for the substrates tested, as compared to the native enzyme. This over-expression protocol now allows for detailed spectroscopic and mechanistic studies on ImiS as well as site-directed mutants of ImiS to be prepared for future structure/function studies. | lld:pubmed |
| pubmed-article:15249050 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:language | eng | lld:pubmed |
| pubmed-article:15249050 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:15249050 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:15249050 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:15249050 | pubmed:month | Aug | lld:pubmed |
| pubmed-article:15249050 | pubmed:issn | 1046-5928 | lld:pubmed |
| pubmed-article:15249050 | pubmed:author | pubmed-author:WalshTimothy... | lld:pubmed |
| pubmed-article:15249050 | pubmed:author | pubmed-author:CrowderMichae... | lld:pubmed |
| pubmed-article:15249050 | pubmed:author | pubmed-author:SpencerJamesJ | lld:pubmed |
| pubmed-article:15249050 | pubmed:author | pubmed-author:CrawfordPatri... | lld:pubmed |
| pubmed-article:15249050 | pubmed:author | pubmed-author:SigdelTaraT | lld:pubmed |
| pubmed-article:15249050 | pubmed:author | pubmed-author:SharmaNarayan... | lld:pubmed |
| pubmed-article:15249050 | pubmed:author | pubmed-author:ChandrasekarS... | lld:pubmed |
| pubmed-article:15249050 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:15249050 | pubmed:volume | 36 | lld:pubmed |
| pubmed-article:15249050 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:15249050 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:15249050 | pubmed:pagination | 272-9 | lld:pubmed |
| pubmed-article:15249050 | pubmed:dateRevised | 2007-11-14 | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:meshHeading | pubmed-meshheading:15249050... | lld:pubmed |
| pubmed-article:15249050 | pubmed:year | 2004 | lld:pubmed |
| pubmed-article:15249050 | pubmed:articleTitle | Over-expression, purification, and characterization of metallo-beta-lactamase ImiS from Aeromonas veronii bv. sobria. | lld:pubmed |
| pubmed-article:15249050 | pubmed:affiliation | Department of Chemistry and Biochemistry, 112 Hughes Hall, Miami University, Oxford, OH 45056, USA. | lld:pubmed |
| pubmed-article:15249050 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:15249050 | pubmed:publicationType | Research Support, U.S. Gov't, P.H.S. | lld:pubmed |
| pubmed-article:15249050 | pubmed:publicationType | Research Support, U.S. Gov't, Non-P.H.S. | lld:pubmed |
| pubmed-article:15249050 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:15249050 | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:15249050 | lld:pubmed |
