Linked Life Data

15649045

Source:http://linkedlifedata.com/resource/pubmed/id/15649045

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2005-1-14
pubmed:abstractText
Chemical cross-linking has proved successful in combination with mass spectrometry as a tool for low-resolution structure determination of proteins. The integration of chemical cross-linking with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry to determine protein interfaces was tested on the calcium-dependent complex between calmodulin (CaM) and a 26-amino acid peptide derived from the skeletal muscle myosin light chain kinase (M13). Different amine-reactive, homobifunctional cross-linkers and a "zero-length" cross-linker were employed. The covalently attached complexes were separated from nonreacted proteins by one-dimensional gel electrophoresis, and the bands of interest were excised and in-gel digested with trypsin. Digestion of the cross-linked complexes resulted in complicated peptide mixtures, which were analyzed by nano-HPLC/nano-ESI-FTICR mass spectrometry. The distance constraints obtained by chemical cross-linking were in agreement with the published NMR structure of the CaM/M13 complex, pointing to residues Lys-18 and Lys-19 of M13 being cross-linked with the central alpha-helix of CaM. Thus, the integrated approach described herein has proven to be an efficient tool for mapping the topology of the CaM/M13 complex. As such it is applicable as a general strategy for the investigation of the spatial organization of protein complexes and complements existing techniques, such as X-ray crystallography and NMR spectroscopy.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0003-2700
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
77
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
495-503
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:15649045-Adipic Acids, pubmed-meshheading:15649045-Calmodulin, pubmed-meshheading:15649045-Chromatography, High Pressure Liquid, pubmed-meshheading:15649045-Cross-Linking Reagents, pubmed-meshheading:15649045-Cyclotrons, pubmed-meshheading:15649045-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:15649045-Ethyldimethylaminopropyl Carbodiimide, pubmed-meshheading:15649045-Fourier Analysis, pubmed-meshheading:15649045-Mass Spectrometry, pubmed-meshheading:15649045-Myosin-Light-Chain Kinase, pubmed-meshheading:15649045-Nanotechnology, pubmed-meshheading:15649045-Nuclear Magnetic Resonance, Biomolecular, pubmed-meshheading:15649045-Peptide Fragments, pubmed-meshheading:15649045-Protein Interaction Mapping, pubmed-meshheading:15649045-Proteins, pubmed-meshheading:15649045-Spectrometry, Mass, Electrospray Ionization, pubmed-meshheading:15649045-Spectrometry, Mass, Matrix-Assisted Laser..., pubmed-meshheading:15649045-Succinates, pubmed-meshheading:15649045-Succinimides
pubmed:year
2005
pubmed:articleTitle
Chemical cross-linking and high-performance Fourier transform ion cyclotron resonance mass spectrometry for protein interaction analysis: application to a calmodulin/target peptide complex.
pubmed:affiliation
Biotechnological-Biomedical Center, Faculty of Chemistry and Mineralogy, University of Leipzig, D-04103 Leipzig, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't