%0 Journal Article
%A Mohrlüder, J.
%A Stangler, T.
%A Wiesehan, K.
%A Hoffmann, Y.
%A Mataruga, A.
%A Willbold, D.
%T Identification of calreticulin as ligand of GABARAP by phage display screening of a peptide library
%J The FEBS journal
%V 274
%N 21
%@ 1742-464X
%C Oxford [u.a.]
%I Wiley-Blackwell
%M PreJuSER-60125
%P 5543 - 5555
%D 2007
%Z Record converted from VDB: 12.11.2012
%X 4-Aminobutyrate type A (GABA(A)) receptor-associated protein (GABARAP) is a ubiquitin-like modifier implicated in the intracellular trafficking of GABA(A) receptors, and belongs to a family of proteins involved in intracellular vesicular transport processes, such as autophagy and intra-Golgi transport. In this article, it is demonstrated that calreticulin is a high affinity ligand of GABARAP. Calreticulin, although best known for its functions as a Ca(2+) -dependent chaperone and a Ca(2+) -buffering protein in the endoplasmic reticulum, is also localized to the cytosol and exerts a variety of extra-endoplasmic reticulum functions. By phage display screening of a randomized peptide library, peptides that specifically bind GABARAP were identified. Their amino acid sequences allowed us to identify calreticulin as a potential GABARAP binding protein. GABARAP binding to calreticulin was confirmed by pull-down experiments with brain lysate and colocalization studies in N2a cells. Calreticulin and GABARAP interact with a dissociation constant K(d) = 64 nm and a mean lifetime of the complex of 20 min. Thus, the interaction between GABARAP and calreticulin is the strongest so far reported for each protein.
%K Amino Acid Motifs
%K Amino Acid Sequence
%K Animals
%K Binding Sites
%K Calreticulin: chemistry
%K Calreticulin: metabolism
%K Cells, Cultured
%K Immunohistochemistry
%K Ligands
%K Microtubule-Associated Proteins: chemistry
%K Microtubule-Associated Proteins: metabolism
%K Models, Molecular
%K Molecular Sequence Data
%K Peptide Fragments: chemistry
%K Peptide Fragments: metabolism
%K Peptide Library
%K Rats
%K Surface Plasmon Resonance
%K Calreticulin (NLM Chemicals)
%K GABARAP protein, rat (NLM Chemicals)
%K Ligands (NLM Chemicals)
%K Microtubule-Associated Proteins (NLM Chemicals)
%K Peptide Fragments (NLM Chemicals)
%K Peptide Library (NLM Chemicals)
%K J (WoSType)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:17916189
%U <Go to ISI:>//WOS:000250297600009
%R 10.1111/j.1742-4658.2007.06073.x
%U https://juser.fz-juelich.de/record/60125