| pubmed-article:9882568 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C0205474 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C0014442 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C0231448 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C0030685 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C0680255 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C0391871 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C1283071 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C1963578 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C1998793 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C1880022 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C1174225 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C0104454 | lld:lifeskim |
| pubmed-article:9882568 | lifeskim:mentions | umls-concept:C1707271 | lld:lifeskim |
| pubmed-article:9882568 | pubmed:issue | 3 | lld:pubmed |
| pubmed-article:9882568 | pubmed:dateCreated | 1999-2-22 | lld:pubmed |
| pubmed-article:9882568 | pubmed:abstractText | Human cytomegalovirus protease (CMV PR) is a target for the development of antiviral therapeutics. To obtain large amounts of native protease, a 268-amino-acid polypeptide with a hexahistidinyl tag at the C terminus was expressed in Escherichia coli. The first 262 amino acids of the recombinant protein were identical to the amino acid sequence of native CMV PR, except for mutations introduced at the internal cleavage site to eliminate autoproteolysis at that site. The hexahistidinyl tag was placed downstream of amino acid 262 of the native CMV PR sequence. In this design, the Ala-Ser bond at amino acids 256-257 constitutes a site naturally cleaved by the protease during capsid maturation. The 268-amino-acid polypeptide with the (His)6 tag was expressed at high levels in E. coli as inclusion bodies. After solubilization of the inclusion bodies, the protease was purified to homogeneity by a single step using Ni2+ affinity chromatography. The protease was refolded to an active enzyme using dialysis which leads to effective autocleavage of the Ala-Ser bond at amino acids 256-257 to remove 12 amino acids including the (His)6 tag from the C terminus of the protein. This strategy yielded large amounts of highly purified CMV PR with the native N terminus and C terminus. Approximately 40 mg of purified CMV PR was obtained per liter of cell culture using this strategy. The enzymatic activity of CMV PR purified from inclusion bodies and refolded to an active enzyme was similar to the enzymatic activity of CMV PR expressed as a soluble protein in E. coli. In addition, the refolded CMV PR could be crystallized for X-ray diffraction. | lld:pubmed |
| pubmed-article:9882568 | pubmed:language | eng | lld:pubmed |
| pubmed-article:9882568 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:9882568 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:9882568 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:9882568 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:9882568 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:9882568 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:9882568 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:9882568 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:9882568 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:9882568 | pubmed:month | Dec | lld:pubmed |
| pubmed-article:9882568 | pubmed:issn | 1046-5928 | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:LullJ MJM | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:BakerC ACA | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:WellsP APA | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:TomasselliA... | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:GarlickR LRL | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:MathewsW RWR | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:TomichC SCS | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:LeoneJ WJW | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:ShellyJ AJA | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:BaldwinE TET | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:FinzelB CBC | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:CurryK AKA | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:PaddockD JDJ | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:MutchlerV TVT | lld:pubmed |
| pubmed-article:9882568 | pubmed:author | pubmed-author:CaveyG SGS | lld:pubmed |
| pubmed-article:9882568 | pubmed:copyrightInfo | Copyright 1998 Academic Press. | lld:pubmed |
| pubmed-article:9882568 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:9882568 | pubmed:volume | 14 | lld:pubmed |
| pubmed-article:9882568 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:9882568 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:9882568 | pubmed:pagination | 343-52 | lld:pubmed |
| pubmed-article:9882568 | pubmed:dateRevised | 2006-11-15 | lld:pubmed |
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| pubmed-article:9882568 | pubmed:meshHeading | pubmed-meshheading:9882568-... | lld:pubmed |
| pubmed-article:9882568 | pubmed:year | 1998 | lld:pubmed |
| pubmed-article:9882568 | pubmed:articleTitle | Recombinant human cytomegalovirus protease with a C-terminal (His)6 extension: purification, autocatalytic release of the mature enzyme, and biochemical characterization. | lld:pubmed |
| pubmed-article:9882568 | pubmed:affiliation | Protein Science, Genomics, Bioprocess Research Preparations, Structural and Analytical and Medicinal Chemistry, Pharmacia & Upjohn, 301 Henrietta Street, Kalamazoo, Michigan, 49007, USA. | lld:pubmed |
| pubmed-article:9882568 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:9882568 | pubmed:publicationType | Comparative Study | lld:pubmed |
