Linked Life Data

10653702

Source:http://linkedlifedata.com/resource/pubmed/id/10653702

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2000-4-3
pubmed:abstractText
Protein function is often regulated by conformational changes that occur in response to ligand binding or covalent modification such as phosphorylation. In many multidomain proteins these conformational changes involve reorientation of domains within the protein. Although X-ray crystallography can be used to determine the relative orientation of domains, the crystal-state conformation can reflect the effect of crystal packing forces and therefore may differ from the physiologically relevant form existing in solution. Here we demonstrate that the solution-state conformation of a multidomain protein can be obtained from its X-ray structure using an extensive set of dipolar couplings measured by triple-resonance multidimensional NMR spectroscopy in weakly aligning solvent. The solution-state conformation of the 370-residue maltodextrin-binding protein (MBP) loaded with beta-cyclodextrin has been determined on the basis of one-bond (15)N-H(N), (15)N-(13)C', (13)C(alpha)-(13)C', two-bond (13)C'-H(N), and three-bond (13)C(alpha)-H(N) dipolar couplings measured for 280, 262, 276, 262, and 276 residues, respectively. This conformation was generated by applying hinge rotations to various X-ray structures of MBP seeking to minimize the difference between the experimentally measured and calculated dipolar couplings. Consistent structures have been derived in this manner starting from four different crystal forms of MBP. The analysis has revealed substantial differences between the resulting solution-state conformation and its crystal-state counterpart (Protein Data Bank accession code 1DMB) with the solution structure characterized by an 11(+/-1) degrees domain closure. We have demonstrated that the precision achieved in these analyses is most likely limited by small uncertainties in the intradomain structure of the protein (ca 5 degrees uncertainty in orientation of internuclear vectors within domains). In addition, potential effects of interdomain motion have been considered using a number of different models and it was found that the structures derived on the basis of dipolar couplings accurately represent the effective average conformation of the protein.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0022-2836
pubmed:author
pubmed:copyrightInfo
Copyright 2000 Academic Press.
pubmed:issnType
Print
pubmed:day
4
pubmed:volume
295
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1265-73
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:10653702-Amino Acid Sequence, pubmed-meshheading:10653702-Bacterial Proteins, pubmed-meshheading:10653702-Carrier Proteins, pubmed-meshheading:10653702-Crystallization, pubmed-meshheading:10653702-Crystallography, X-Ray, pubmed-meshheading:10653702-Cyclodextrins, pubmed-meshheading:10653702-Escherichia coli, pubmed-meshheading:10653702-Escherichia coli Proteins, pubmed-meshheading:10653702-Models, Molecular, pubmed-meshheading:10653702-Molecular Sequence Data, pubmed-meshheading:10653702-Motion, pubmed-meshheading:10653702-Nuclear Magnetic Resonance, Biomolecular, pubmed-meshheading:10653702-Protein Structure, Tertiary, pubmed-meshheading:10653702-Reproducibility of Results, pubmed-meshheading:10653702-Sensitivity and Specificity, pubmed-meshheading:10653702-Solutions, pubmed-meshheading:10653702-Solvents, pubmed-meshheading:10653702-beta-Cyclodextrins
pubmed:year
2000
pubmed:articleTitle
Orienting domains in proteins using dipolar couplings measured by liquid-state NMR: differences in solution and crystal forms of maltodextrin binding protein loaded with beta-cyclodextrin.
pubmed:affiliation
Contribution Protein Engineering Network Centres of Excellence and Department of Medical Genetics, University of Toronto, Toronto, Ontario, M5S 1A8, Canada. nikolai@pound.med.utoronto.ca
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't