Document Detail

Noxa/Bcl-2 protein interactions contribute to bortezomib resistance in human lymphoid cells.
MedLine Citation:
PMID:  21454712     Owner:  NLM     Status:  MEDLINE    
Previous studies have suggested that the BH3 domain of the proapoptotic Bcl-2 family member Noxa only interacts with the anti-apoptotic proteins Mcl-1 and A1 but not Bcl-2. In view of the similarity of the BH3 binding domains of these anti-apoptotic proteins as well as recent evidence that studies of isolated BH3 domains can potentially underestimate the binding between full-length Bcl-2 family members, we examined the interaction of full-length human Noxa with anti-apoptotic human Bcl-2 family members. Surface plasmon resonance using bacterially expressed proteins demonstrated that Noxa binds with mean dissociation constants (K(D)) of 3.4 nm for Mcl-1, 70 nm for Bcl-x(L), and 250 nm for wild type human Bcl-2, demonstrating selectivity but not absolute specificity of Noxa for Mcl-1. Further analysis showed that the Noxa/Bcl-2 interaction reflected binding between the Noxa BH3 domain and the Bcl-2 BH3 binding groove. Analysis of proteins expressed in vivo demonstrated that Noxa and Bcl-2 can be pulled down together from a variety of cells. Moreover, when compared with wild type Bcl-2, certain lymphoma-derived Bcl-2 mutants bound Noxa up to 20-fold more tightly in vitro, pulled down more Noxa from cells, and protected cells against killing by transfected Noxa to a greater extent. When killing by bortezomib (an agent whose cytotoxicity in Jurkat T-cell leukemia cells is dependent on Noxa) was examined, apoptosis was enhanced by the Bcl-2/Bcl-x(L) antagonist ABT-737 or by Bcl-2 down-regulation and diminished by Bcl-2 overexpression. Collectively, these observations not only establish the ability of Noxa and Bcl-2 to interact but also identify Bcl-2 overexpression as a potential mechanism of bortezomib resistance.
Alyson J Smith; Haiming Dai; Cristina Correia; Rie Takahashi; Sun-Hee Lee; Ingo Schmitz; Scott H Kaufmann
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2011-03-22
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  286     ISSN:  1083-351X     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2011 May 
Date Detail:
Created Date:  2011-05-16     Completed Date:  2011-07-26     Revised Date:  2014-04-23    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  17682-92     Citation Subset:  IM    
Copyright Information:
© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.
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MeSH Terms
Antineoplastic Agents / pharmacology*
Apoptosis / drug effects
Boronic Acids / pharmacology*
Drug Resistance, Neoplasm / drug effects*
Hematologic Neoplasms / genetics,  metabolism
Jurkat Cells
Lymphocytes / metabolism*
Myeloid Cell Leukemia Sequence 1 Protein
Protein Binding
Protein Structure, Tertiary
Proto-Oncogene Proteins c-bcl-2 / genetics,  metabolism*
Pyrazines / pharmacology*
Grant Support
Reg. No./Substance:
0/Antineoplastic Agents; 0/Boronic Acids; 0/Myeloid Cell Leukemia Sequence 1 Protein; 0/PMAIP1 protein, human; 0/Proto-Oncogene Proteins c-bcl-2; 0/Pyrazines; 0/bortezomib

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