Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here for more information

Click here to sign up for SAGE Journal Email Alerts today!

Sign In to gain access to subscriptions and/or personal tools.
Journal of Biomolecular Screening
This Article
Right arrow Full Text (PDF)
Right arrow References
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Veniere, S.
Right arrow Articles by Lambrechts, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Veniere, S.
Right arrow Articles by Lambrechts, A.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

The Interaction of Proline-Rich Ligands with Profilin Probed with an Enzyme-Linked Immunosorbent Assay

Sylvie Veniere

Department of Medical Protein Research, VIB, Ghent, Belgium, Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium

Christophe Ampe

Department of Medical Protein Research, VIB, Ghent, Belgium, Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium

Joël Vandekerckhove

Department of Medical Protein Research, VIB, Ghent, Belgium, Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium

Anja Lambrechts

Department of Medical Protein Research, VIB, Ghent, Belgium, Anja.lambrechts{at}ugent.be, Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium

To detect interactions of different proline-rich ligands with profilins, the authors developed a simple analytical antibody-based screening method. Profilin I or profilin IIa was coated in microplates, and ligand binding was monitored via antibody detection. Using purified components, the authors show that the assay is very sensitive as nanomolar concentrations of recombinant profilin ligands can be used. They further apply this technique to detect interaction of profilin with various proline-rich partners, either endogenously present or ectopically expressed as tagged fusions, using lysates. With this assay, the authors identify Shootin1 as a novel profilin IIa partner. In addition, they demonstrate that this assay can be used for studying competition or ternary complex formation. In conclusion, they developed a sensitive, easy-to-use, and versatile method for the study of the interaction between profilin and different ligands. (Journal of Biomolecular Screening 2009:350-359)

Key Words: profilin isoforms • poly(L-proline) • Ena/VASP proteins • mDia • SH3 domain • Shootin1

Journal of Biomolecular Screening, Vol. 14, No. 4, 350-359 (2009)
DOI: 10.1177/1087057109332594
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?