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Analysis of the adaptor function of the LIM domain-containing protein FHL2 using an affinity chromatography approach

✍ Scribed by Haquima El Mourabit; Stefan Müller; Lucy Tunggal; Mats Paulsson; Dr. Monique Aumailley

Publisher
John Wiley and Sons
Year
2004
Tongue
English
Weight
655 KB
Volume
92
Category
Article
ISSN
0730-2312

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✦ Synopsis

Abstract

Containing four LIM domains and an N‐terminal half LIM domain, FHL2 has been predicted to have an adaptor function in the formation of higher order molecular complexes in the nucleus and the cytoplasm of cells. We expressed recombinant FHL2 in insect cells using the baculovirus system and used it to isolate direct or indirect interaction partners from the cytosolic fraction of fibroblasts by affinity chromatography. These were identified by their peptide mass fingerprints using MALDI‐TOF mass spectrometry. Cytoskeleton‐associated proteins present among the bound proteins were shown to co‐localise with FHL2 in cell lamellipodia by indirect immunofluorescence staining. © 2004 Wiley‐Liss, Inc.

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