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Open Access Highly Accessed Research article

Bcr is a substrate for Transglutaminase 2 cross-linking activity

Sun-Ju Yi1, John Groffen123 and Nora Heisterkamp123*

Author affiliations

1 Section of Molecular Carcinogenesis, Division of Hematology/Oncology, Ms#54, Childrens Hospital Los Angeles, 4650 Sunset Boulevard, Los Angeles CA 90027, USA

2 The Saban Research Institute of Childrens Hospital Los Angeles, CA 90027, USA

3 Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA

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Citation and License

BMC Biochemistry 2011, 12:8  doi:10.1186/1471-2091-12-8

Published: 10 February 2011

Abstract

Background

Breakpoint cluster region (Bcr) is a multi-domain protein that contains a C-terminal GTPase activating protein (GAP) domain for Rac. Transglutaminase 2 (TG2) regulates Bcr by direct binding to its GAP domain. Since TG2 has transglutaminase activity that has been implicated in the response to extreme stress, we investigated if Bcr can also act as a substrate for TG2.

Results

We here report that activation of TG2 by calcium caused the formation of covalently cross-linked Bcr. Abr, a protein related to Bcr but lacking its N-terminal oligomerization domain, was not cross-linked by TG2 even though it forms a complex with it. A Bcr mutant missing the first 62 amino acid residues remained monomeric in the presence of activated TG2, showing that this specific domain is necessary for the cross-linking reaction. Calcium influx induced by a calcium ionophore in primary human endothelial cells caused cross-linking of endogenous Bcr, which was inhibited by the TG2 inhibitor cystamine. Treatment of cells with cobalt chloride, a hypoxia-mimetic that causes cellular stress, also generated high molecular weight Bcr complexes. Cross-linked Bcr protein appeared in the TritonX-100-insoluble cell fraction and further accumulated in cells treated with a proteasome inhibitor.

Conclusions

Bcr thus represents both an interacting partner under non-stressed conditions and a target of transglutaminase activity for TG2 during extreme stress.