Document Detail

Identification of a novel 14-3-3zeta binding site within the cytoplasmic tail of platelet glycoprotein Ibalpha.
MedLine Citation:
PMID:  15054037     Owner:  NLM     Status:  MEDLINE    
The glycoprotein Ib-V-IX (GPIb-V-IX) complex interacts with subendothelial von Willebrand factor (VWF) to ensure recruitment of platelets at sites of vascular injury, a process that culminates in integrin alpha(IIb)beta(3)-dependent stable adhesion and spreading. Interaction of the 14-3-3zeta adaptor protein with the C-terminal 606-610 phosphoserine motif of the GPIbalpha subunit has been implicated in the control of alpha(IIb)beta(3) activation and cell spreading. In this study, we have examined potentially novel 14-3-3zeta binding sites by expressing mutant forms of GPIbalpha in Chinese-hamster-ovary (CHO) cells. Analysis of a series of neighboring 11-12 residue deletions identified a critical role for the 580-LVAGRRPSALS-590 sequence in promoting GPIbalpha-14-3-3zeta interaction. Development of a phosphospecific antibody demonstrated high levels of phosphorylation of the Ser587 and Ser590 residues in resting platelets (which became dephosphorylated during platelet spreading on VWF), and peptides containing these phosphorylated residues effectively displaced 14-3-3zeta from GPIbalpha. Analysis of single and double alanine substitutions of Ser587 and Ser590 demonstrated a major role for these residues in promoting GPIbalpha-14-3-3zeta binding. Moreover, these cell lines exhibited a defect in cell spreading on immobilized VWF. These studies demonstrate the existence of a second major 14-3-3zeta binding site within the cytoplasmic tail of GPIbalpha that has an important functional role in regulating integrin-dependent cell spreading.
Pierre Mangin; Tovo David; Vincent Lavaud; Susan L Cranmer; Inna Pikovski; Shaun P Jackson; Michael C Berndt; Jean-Pierre Cazenave; Christian Gachet; François Lanza
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2004-03-30
Journal Detail:
Title:  Blood     Volume:  104     ISSN:  0006-4971     ISO Abbreviation:  Blood     Publication Date:  2004 Jul 
Date Detail:
Created Date:  2004-07-02     Completed Date:  2004-08-17     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7603509     Medline TA:  Blood     Country:  United States    
Other Details:
Languages:  eng     Pagination:  420-7     Citation Subset:  AIM; IM    
INSERM U.311, Etablissement Français du Sang-Alsace, 10 rue Spielmann, BP 36, 67065 Strasbourg Cedex, France.
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MeSH Terms
14-3-3 Proteins
Amino Acid Substitution
Binding Sites
Blood Platelets / cytology,  metabolism*
CHO Cells
Cytoplasm / metabolism
Gene Deletion
Integrins / metabolism
Platelet Glycoprotein GPIb-IX Complex / chemistry,  genetics*,  metabolism*
Protein Structure, Tertiary
Serine / metabolism
Tyrosine 3-Monooxygenase / metabolism*
von Willebrand Factor / metabolism,  pharmacology
Grant Support
R01 HL 50744-02/HL/NHLBI NIH HHS
Reg. No./Substance:
0/14-3-3 Proteins; 0/Integrins; 0/Platelet Glycoprotein GPIb-IX Complex; 0/von Willebrand Factor; 56-45-1/Serine; EC 3-Monooxygenase

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