| pubmed-article:16750914 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0033684 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0022023 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0030956 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0439855 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0037813 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0231881 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0010600 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0282183 | lld:lifeskim |
| pubmed-article:16750914 | lifeskim:mentions | umls-concept:C0204514 | lld:lifeskim |
| pubmed-article:16750914 | pubmed:issue | 8 | lld:pubmed |
| pubmed-article:16750914 | pubmed:dateCreated | 2006-7-31 | lld:pubmed |
| pubmed-article:16750914 | pubmed:abstractText | For structural studies of proteins and their complexes, chemical cross-linking combined with mass spectrometry presents a promising strategy to obtain structural data of protein interfaces from low quantities of proteins within a short time. We explore the use of isotope-labeled cross-linkers in combination with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry for a more efficient identification of cross-linker containing species. For our studies, we chose the calcium-independent complex between calmodulin and a 25-amino acid peptide from the C-terminal region of adenylyl cyclase 8 containing an "IQ-like motif." Cross-linking reactions between calmodulin and the peptide were performed in the absence of calcium using the amine-reactive, isotope-labeled (d0 and d4) cross-linkers BS3 (bis[sulfosuccinimidyl]suberate) and BS2G (bis[sulfosuccinimidyl]glutarate). Tryptic in-gel digestion of excised gel bands from covalently cross-linked complexes resulted in complicated peptide mixtures, which were analyzed by nano-HPLC/nano-ESI-FTICR mass spectrometry. In cases where more than one reactive functional group, e.g., amine groups of lysine residues, is present in a sequence stretch, MS/MS analysis is a prerequisite for unambiguously identifying the modified residues. MS/MS experiments revealed two lysine residues in the central alpha-helix of calmodulin as well as three lysine residues both in the C-terminal and N-terminal lobes of calmodulin to be cross-linked with one single lysine residue of the adenylyl cyclase 8 peptide. Further cross-linking studies will have to be conducted to propose a structural model for the calmodulin/peptide complex, which is formed in the absence of calcium. The combination of using isotope-labeled cross-linkers, determining the accurate mass of intact cross-linked products, and verifying the amino acid sequences of cross-linked species by MS/MS presents a convenient approach that offers the perspective to obtain structural data of protein assemblies within a few days. | lld:pubmed |
| pubmed-article:16750914 | pubmed:language | eng | lld:pubmed |
| pubmed-article:16750914 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:16750914 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:16750914 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:16750914 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:16750914 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:16750914 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:16750914 | pubmed:month | Aug | lld:pubmed |
| pubmed-article:16750914 | pubmed:issn | 1044-0305 | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:CooperDermot... | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:Beck-Sickinge... | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:SchmidtAndrea... | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:IhlingChristi... | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:MechtlerKarlK | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:SinzAndreaA | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:HaackMichaelM | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:SchulzDaniela... | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:StinglChristo... | lld:pubmed |
| pubmed-article:16750914 | pubmed:author | pubmed-author:KalkhofStefan... | lld:pubmed |
| pubmed-article:16750914 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:16750914 | pubmed:volume | 17 | lld:pubmed |
| pubmed-article:16750914 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:16750914 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:16750914 | pubmed:pagination | 1100-13 | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
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| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:meshHeading | pubmed-meshheading:16750914... | lld:pubmed |
| pubmed-article:16750914 | pubmed:year | 2006 | lld:pubmed |
| pubmed-article:16750914 | pubmed:articleTitle | Isotope-labeled cross-linkers and Fourier transform ion cyclotron resonance mass spectrometry for structural analysis of a protein/peptide complex. | lld:pubmed |
| pubmed-article:16750914 | pubmed:affiliation | Biotechnological-Biomedical Center, Faculty of Chemistry and Mineralogy, University of Leipzig, Leipzig, Germany. | lld:pubmed |
| pubmed-article:16750914 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:16750914 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
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