Linked Life Data

11170476

Source:http://linkedlifedata.com/resource/pubmed/id/11170476

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2001-2-22
pubmed:abstractText
Animals vaccinated with heat shock protein (HSP)--peptide complexes develop specific protective immunity against cancers from which the HSPs were originally isolated. This autologous specific immunity has been demonstrated using a number of HSP--peptide antigen complexes. A prototypical HSP-based cancer vaccine is the gp96--peptide antigen complex, which is currently undergoing human clinical trials. Here, we analyzed the structure of a recombinant wild-type and a mutant gp96 protein and their peptide complexes using a number of biophysical techniques. Gel filtration chromatography, dynamic light scattering, and equilibrium analytical ultracentrifugation demonstrated that both a wild-type gp96 and a gp96 mutant lacking a dimerization domain formed higher order structures. More detailed analysis using scanning transmission electron microscopy indicated that both the wild-type and dimerization deletion mutant gp96 protein were organized, unexpectedly, into large aggregates. Size distributions ranged from dimers to octamers and higher. Circular dichroism and intrinsic Trp fluorescence suggested that the gp96 dimerization domain deletion mutant protein was more compact than the wild-type gp96. A fluorescent peptide antigen was synthesized, and the peptide-binding properties of wild-type and the dimerization domain deletion mutant gp96 were studied. Fluorescence lifetime and anisotropy decay showed that the bound antigenic peptide was located in a hydrophobic pocket, with considerable free space for the rotation of the probe. Deletion of the dimerization domain affected the peptide-binding microenvironment, although peptide-binding affinity was reduced by only a small extent. Peptide--gp96 complexes were extremely stable, persisting for many days in the cold. The extraordinary stability of peptide--gp96 complexes and the plasticity of the peptide-binding pocket support the proposed relay of diverse peptides to MHC and/or other molecules via molecular recognition.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
6
pubmed:volume
40
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1483-95
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:11170476-Animals, pubmed-meshheading:11170476-Antigen Presentation, pubmed-meshheading:11170476-Antigens, Neoplasm, pubmed-meshheading:11170476-Chromatography, Affinity, pubmed-meshheading:11170476-Chromatography, High Pressure Liquid, pubmed-meshheading:11170476-Circular Dichroism, pubmed-meshheading:11170476-Drug Stability, pubmed-meshheading:11170476-Endoplasmic Reticulum, pubmed-meshheading:11170476-Fluorescence Polarization, pubmed-meshheading:11170476-HSP70 Heat-Shock Proteins, pubmed-meshheading:11170476-Heat-Shock Proteins, pubmed-meshheading:11170476-Light, pubmed-meshheading:11170476-Macromolecular Substances, pubmed-meshheading:11170476-Membrane Proteins, pubmed-meshheading:11170476-Mice, pubmed-meshheading:11170476-Mutagenesis, Site-Directed, pubmed-meshheading:11170476-Peptides, pubmed-meshheading:11170476-Protein Binding, pubmed-meshheading:11170476-Scattering, Radiation, pubmed-meshheading:11170476-Spectrometry, Fluorescence, pubmed-meshheading:11170476-Spectrometry, Mass, Matrix-Assisted Laser..., pubmed-meshheading:11170476-Ultracentrifugation
pubmed:year
2001
pubmed:articleTitle
Biophysical analysis of the endoplasmic reticulum-resident chaperone/heat shock protein gp96/GRP94 and its complex with peptide antigen.
pubmed:affiliation
Laboratory of Molecular Genetics, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't