Linked Life Data

11705363

Source:http://linkedlifedata.com/resource/pubmed/id/11705363

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
46
pubmed:dateCreated
2001-11-13
pubmed:abstractText
Using peptides tethered to polymer microbeads, we have developed a technique for measuring the interactions between the transmembrane alpha-helices of membrane proteins and for screening combinatorial libraries of peptides for members that interact with specific helices from membrane proteins. The method was developed using the well-characterized homodimerization sequence of the membrane-spanning alpha-helix from the erythrocyte membrane protein glycophorin A (GPA). As a control, we also tested a variant with a dimer-disrupting alteration of a critical glycine residue to leucine. To test for detectable, native interactions between detergent-solubilized and microbead-tethered alpha-helices, we incubated fluorescent dye-labeled GPA analogues in sodium dodecyl sulfate solution with microbeads that contained covalently attached GPA analogues. When the dye-labeled peptide in solution and the bead-tethered peptide both contained the native glycophorin A sequence, the microbeads readily accumulated the dye through lateral peptide-peptide interactions and were visibly fluorescent under UV light. When either the peptide in solution or the peptide attached to the beads contained the glycine to leucine change, the beads did not accumulate any dye. The usefulness of this method for screening tethered peptide libraries was tested by incubating dye-labeled, native sequence peptides in detergent solution with a few native sequence beads plus an excess of beads containing the variant glycine to leucine sequence. When the dye-labeled peptide in solution was present at a concentration of > or =2 microM, the few native sequence beads were visually distinguishable from the others because of their bright fluorescence. Using this model system, we have shown that it is possible to visually detect specific, native interactions between alpha-helices from membrane proteins using peptides tethered to polymer microbeads. It will thus be possible to use this method to measure the specific lateral interactions that drive the folding and organization of membrane proteins and to screen combinatorial libraries of peptides for members that interact with them.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
20
pubmed:volume
40
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
13753-9
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:11705363-Amino Acid Sequence, pubmed-meshheading:11705363-Dimerization, pubmed-meshheading:11705363-Fluorescent Dyes, pubmed-meshheading:11705363-Glycophorin, pubmed-meshheading:11705363-Mass Spectrometry, pubmed-meshheading:11705363-Membrane Proteins, pubmed-meshheading:11705363-Micelles, pubmed-meshheading:11705363-Microspheres, pubmed-meshheading:11705363-Models, Molecular, pubmed-meshheading:11705363-Molecular Sequence Data, pubmed-meshheading:11705363-Peptides, pubmed-meshheading:11705363-Polymers, pubmed-meshheading:11705363-Polystyrenes, pubmed-meshheading:11705363-Protein Structure, Secondary, pubmed-meshheading:11705363-Solubility, pubmed-meshheading:11705363-Solutions, pubmed-meshheading:11705363-Spectrometry, Fluorescence, pubmed-meshheading:11705363-Xanthenes
pubmed:year
2001
pubmed:articleTitle
Visual detection of specific, native interactions between soluble and microbead-tethered alpha-helices from membrane proteins.
pubmed:affiliation
Department of Biochemistry, SL43, Tulane University Health Sciences Center, New Orleans, Louisiana 70112-2699, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't