Linked Life Data

12628256

Source:http://linkedlifedata.com/resource/pubmed/id/12628256

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2003-3-11
pubmed:abstractText
Topological linking of proteins is a new approach for stabilizing and controlling the oligomerization state of proteins that fold in an interwined manner. The recent design of a backbone cyclized protein catenane based on the p53tet domain suggested that topological cross-linking provided increased stability against thermal and chemical denaturation. However, the tetrameric structure complicated detailed biophysical analysis of this protein. Here, we describe the design, synthesis and thermodynamic characterization of a protein catenane based on a dimeric mutant of the p53tet domain (M340E/L344K). The formation of the catenane proceeded efficiently, and the overall structure and oligomerization of the domain was not affected by the formation of the topological link. Unfolding and refolding of the catenane was consistent with a two-state process. The topological link stabilized the dimer against thermal and chemical denaturation considerably, raising the apparent melting temperature by 59 degrees C and the midpoint of denaturation by 4.5M GuHCl at a concentration of 50 microM. The formation of the topological link increased the resistance of the dimer to proteolysis. However, the m value decreased by 1.7kcalmol(-1)M(-1), suggesting a decrease in accessible surface area in the unfolded state. This implies that the stabilization from the topological link is largely due to a destabilization of the unfolded state, similar to other cross-links in proteins. Topological linking therefore provides a powerful and orthogonal tool for the stabilization of peptide and protein oligomers.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0022-2836
pubmed:author
pubmed:issnType
Print
pubmed:day
21
pubmed:volume
327
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
537-48
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:12628256-Anthracenes, pubmed-meshheading:12628256-Chromatography, High Pressure Liquid, pubmed-meshheading:12628256-Circular Dichroism, pubmed-meshheading:12628256-Cross-Linking Reagents, pubmed-meshheading:12628256-Dimerization, pubmed-meshheading:12628256-Disulfides, pubmed-meshheading:12628256-Guanidine, pubmed-meshheading:12628256-Humans, pubmed-meshheading:12628256-Mass Spectrometry, pubmed-meshheading:12628256-Models, Molecular, pubmed-meshheading:12628256-Peptide Fragments, pubmed-meshheading:12628256-Peptide Hydrolases, pubmed-meshheading:12628256-Polycyclic Compounds, pubmed-meshheading:12628256-Protein Conformation, pubmed-meshheading:12628256-Protein Denaturation, pubmed-meshheading:12628256-Protein Engineering, pubmed-meshheading:12628256-Protein Folding, pubmed-meshheading:12628256-Spectrometry, Fluorescence, pubmed-meshheading:12628256-Thermodynamics, pubmed-meshheading:12628256-Tumor Suppressor Protein p53, pubmed-meshheading:12628256-Ultracentrifugation
pubmed:year
2003
pubmed:articleTitle
Thermodynamics of a designed protein catenane.
pubmed:affiliation
Department of Cell Biology and Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, MC CVN-6, La Jolla, CA 92037, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't