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Zinc binding site in PICK1 is dominantly located at the CPC motif of its PDZ domain
Author(s) -
Shi Yawei,
Zhang Lei,
Yuan Jingming,
Xiao Hong,
Yang Xiuqing,
Niu Lixi
Publication year - 2008
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2008.05434.x
Subject(s) - pdz domain , binding site , chemistry , mutant , signal transducing adaptor protein , binding domain , lim domain , plasma protein binding , zinc finger , c terminus , cytoplasm , scaffold protein , biology , biophysics , biochemistry , microbiology and biotechnology , amino acid , phosphorylation , transcription factor , signal transduction , gene
PICK1 ( p rotein i nteracting with C k inase 1) containing a PDZ domain, a BAR domain, and two short acidic regions is as an adaptor protein that plays an important role in α‐amino‐3‐hydroxy‐5‐methylisoxazole‐4‐propionic acid receptor trafficking, cell morphology and migration, as well as in some diseases such as cancer, schizophrenia and pain. To better understand the physiological function of PICK1, we expressed the recombinant PICK1 and its truncated mutants in E.coli, and measured their zinc binding properties by fluorescence and competition assay. It is shown that PICK1 has one Zn 2+ ‐binding site. The Zn 2+ ‐binding properties of PICK1 are not appreciably affected after the removal of BARC domain (involving BAR domain and C‐terminal acidic region). Deleting the N‐terminal acidic region of NPDZ domain (involving PDZ domain and N‐terminal acidic region) in PICK1 impairs its Zn 2+ ‐binding capacity.The mutation of the CPC (Cys‐Pro‐Cys) motif in the PDZ domain of PICK1 abolishes the ability of Zn 2+ ‐binding. In addition, Zn 2+ can enhance the lipid‐binding ability of PDZ domain as observed in both protein‐lipid overlay assay and fluorescence analysis. The results presented in this report suggested that Zn 2+ plays a regulatory role in the trafficking of PICK1 from the cytoplasm to cell membrane.