| pubmed-article:19301314 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C0002520 | lld:lifeskim |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C2936473 | lld:lifeskim |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C1511790 | lld:lifeskim |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C1517880 | lld:lifeskim |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C0243126 | lld:lifeskim |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C0597717 | lld:lifeskim |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C0016315 | lld:lifeskim |
| pubmed-article:19301314 | lifeskim:mentions | umls-concept:C0303920 | lld:lifeskim |
| pubmed-article:19301314 | pubmed:issue | 6 | lld:pubmed |
| pubmed-article:19301314 | pubmed:dateCreated | 2009-4-13 | lld:pubmed |
| pubmed-article:19301314 | pubmed:abstractText | Position-specific incorporation of fluorescent groups is a useful method for analysis of the functions and structures of proteins. We have developed a method for the incorporation of visible-wavelength-fluorescent non-natural amino acids into proteins in a cell-free translation system. Using this technique, we introduced one or two BODIPY-linked amino acids into maltose-binding protein (MBP) to obtain MBP derivatives showing ligand-dependent changes in fluorescence intensity or intensity ratio. BODIPY-FL-aminophenylalanine was incorporated in place of 15 tyrosines, as well as the N-terminal Lys1, and the C-terminal Lys370 of MBP. Fluorescence measurements revealed that MBP containing a BODIPY-FL moiety in place of Tyr210 showed a 13-fold increase in fluorescence upon binding of maltose. Tryptophan-to-phenylalanine substitutions suggest that the increase in fluorescence was the result of a decrease in the quenching of BODIPY-FL by tryptophan located around the binding site. MBP containing a BODIPY-558 moiety also showed a maltose-dependent increase in fluorescence. BODIPY-FL was then additionally incorporated in place of Lys1 of the BODIPY-558-containing MBP as a response to the amber codon. Fluorescence measurements with excitation of BODIPY-FL showed a large change in fluorescence intensity ratio (0.13 to 1.25) upon binding of maltose; this change can be attributed to fluorescence resonance energy transfer (FRET) and maltose-dependent quenching of BODIPY-558. These results demonstrate the usefulness of the position-specific incorporation of fluorescent amino acids in the fluorescence-based detection of protein functions. | lld:pubmed |
| pubmed-article:19301314 | pubmed:language | eng | lld:pubmed |
| pubmed-article:19301314 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19301314 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:19301314 | pubmed:month | Apr | lld:pubmed |
| pubmed-article:19301314 | pubmed:issn | 1439-7633 | lld:pubmed |
| pubmed-article:19301314 | pubmed:author | pubmed-author:HohsakaTakahi... | lld:pubmed |
| pubmed-article:19301314 | pubmed:author | pubmed-author:IijimaIsseiI | lld:pubmed |
| pubmed-article:19301314 | pubmed:issnType | Electronic | lld:pubmed |
| pubmed-article:19301314 | pubmed:day | 17 | lld:pubmed |
| pubmed-article:19301314 | pubmed:volume | 10 | lld:pubmed |
| pubmed-article:19301314 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:19301314 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:19301314 | pubmed:pagination | 999-1006 | lld:pubmed |
| pubmed-article:19301314 | pubmed:dateRevised | 2010-11-18 | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:meshHeading | pubmed-meshheading:19301314... | lld:pubmed |
| pubmed-article:19301314 | pubmed:year | 2009 | lld:pubmed |
| pubmed-article:19301314 | pubmed:articleTitle | Position-specific incorporation of fluorescent non-natural amino acids into maltose-binding protein for detection of ligand binding by FRET and fluorescence quenching. | lld:pubmed |
| pubmed-article:19301314 | pubmed:affiliation | Japan Advanced Institute of Science and Technology, Asahidai, Nomi, Ishikawa, Japan. | lld:pubmed |
| pubmed-article:19301314 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:19301314 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
