Linked Life Data

11373085

Source:http://linkedlifedata.com/resource/pubmed/id/11373085

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2001-5-24
pubmed:abstractText
Polypeptides corresponding to hydrophobic transmembrane alpha-helices, such as residues 69-101 of glycophorin A, are notoriously difficult to prepare in quantities sufficient for biophysical experiments. Simple synthetic and purification approaches reported here have been developed by combining a few modifications to standard procedures, without resorting to elevated temperatures, expensive activation strategies, or complex hydrophobic solvent mixtures. The cost of screening projects, preparing labeled peptides, and examining sequence variations is thereby significantly reduced. The quality of the peptide synthesized by this small-scale 9-fluorenylmethoxycarbonyl (Fmoc) strategy is comparable to that of the peptide synthesized by an experienced resource facility using a large-scale tert-butyloxycarbonyl strategy. Using reverse-phase HPLC, the desired peptide was separated from the primary side product (a Leu or Ile deletion) and quantitatively recovered at greater than 98% purity. Baseline resolution was achieved using a water:acetonitrile gradient to elute the peptides from a cyanopropyl column at ambient temperature. Combining these approaches readily yields 10 to 20 mg of pure transmembrane peptide from a small-scale Fmoc synthesis. The approaches are readily transferable to transmembrane sequences not previously synthesized and do not require setting up a specialized facility. The time and start-up expense required to launch new studies are thereby reduced expanding the range and detail with which questions in membrane protein biophysics can be explored.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0003-2697
pubmed:author
pubmed:copyrightInfo
Copyright 2001 Academic Press.
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
293
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
102-8
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
2001
pubmed:articleTitle
High-yield synthesis and purification of an alpha-helical transmembrane domain.
pubmed:affiliation
Department of Chemistry, Yale University, New Haven, CT 06520, 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