A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells

Saathoff, Hinnerk, Brofelth, Mattias, Trinh, Anne, Parker, Benjamin L., Ryan, Daniel P., Low, Jason K. K., Webb, Sarah R., Silva, Ana P. G., Mackay, Joel P. and Shepherd, Nicholas E. (2015) A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells. Bioorganic and Medicinal Chemistry, 23 5: 960-965. doi:10.1016/j.bmc.2015.01.023


Author Saathoff, Hinnerk
Brofelth, Mattias
Trinh, Anne
Parker, Benjamin L.
Ryan, Daniel P.
Low, Jason K. K.
Webb, Sarah R.
Silva, Ana P. G.
Mackay, Joel P.
Shepherd, Nicholas E.
Title A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells
Journal name Bioorganic and Medicinal Chemistry   Check publisher's open access policy
ISSN 1464-3391
0968-0896
Publication date 2015-03-01
Sub-type Article (original research)
DOI 10.1016/j.bmc.2015.01.023
Volume 23
Issue 5
Start page 960
End page 965
Total pages 6
Place of publication Kidlington, Oxford, United Kingdom
Publisher Pergamon Press
Collection year 2016
Language eng
Formatted abstract
We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1–15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST–FOG1(1–45).
Keyword Affinity purification
Friend of GATA1 (FOG1)
Multi-protein complex
Nucleosome remodeling and deacetylase complex peptides
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Non HERDC
Institute for Molecular Bioscience - Publications
 
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