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Thioflavin S (NSC71948) interferes with Bcl-2-associated athanogene (BAG-1)-mediated protein-protein interactions

Thioflavin S (NSC71948) interferes with Bcl-2-associated athanogene (BAG-1)-mediated protein-protein interactions
Thioflavin S (NSC71948) interferes with Bcl-2-associated athanogene (BAG-1)-mediated protein-protein interactions
The C-terminal BAG domain is thought to play a key role in BAG-1-induced survival and proliferation by mediating protein-protein interactions, for example, with heat shock proteins HSC70 and HSP70, and with RAF-1 kinase. Here, we have identified thioflavin S (NSC71948) as a potential small-molecule chemical inhibitor of these interactions. NSC71948 inhibited the interaction of BAG-1 and HSC70 in vitro and decreased BAG-1:HSC70 and BAG-1:HSP70 binding in intact cells. NSC71948 also reduced binding between BAG-1 and RAF-1, but had no effect on the interaction between two unrelated proteins, BIM and MCL-1. NSC71948 functionally reversed the ability of BAG-1 to promote vitamin D3 receptor-mediated transactivation, an activity of BAG-1 that depends on HSC70/HSP70 binding, and reduced phosphorylation of p44/42 mitogen-activate protein kinase. NSC71948 can be used to stain amyloid fibrils; however, structurally related compounds, thioflavin T and BTA-1, had no effect on BAG-1:HSC70 binding, suggesting that structural features important for amyloid fibril binding and inhibition of BAG-1:HSC70 binding may be separable. We demonstrated that NSC71948 inhibited the growth of BAG-1 expressing human ZR-75-1 breast cancer cells and wild-type, but not BAG-1-deficient, mouse embryo fibroblasts. Taken together, these data suggest that NSC71948 may be a useful molecule to investigate the functional significance of BAG-1 C-terminal protein interactions. However, it is important to recognize that NSC71948 may exert additional “off-target” effects. Inhibition of BAG-1 function may be an attractive strategy to inhibit the growth of BAG-1-overexpressing cancers, and further screens of additional compound collections may be warranted
0022-3565
680-689
Sharp, Adam
f26a1894-b10f-4864-a314-750628d6c0cc
Crabb, Simon J.
bcd1b566-7677-4f81-8429-3ab0e85f8373
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
Hague, Angela
0e1199e4-08c3-4e8b-b40f-2fc61555e6f4
Cutress, Ramsey
68ae4f86-e8cf-411f-a335-cdba51797406
Townsend, Paul A.
a2680443-664e-46d0-b4dd-97456ba810db
Ganesan, A.
62aa5a87-9308-4383-8686-99726b6bcfb9
Packham, Graham
fdabe56f-2c58-469c-aadf-38878f233394
Sharp, Adam
f26a1894-b10f-4864-a314-750628d6c0cc
Crabb, Simon J.
bcd1b566-7677-4f81-8429-3ab0e85f8373
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
Hague, Angela
0e1199e4-08c3-4e8b-b40f-2fc61555e6f4
Cutress, Ramsey
68ae4f86-e8cf-411f-a335-cdba51797406
Townsend, Paul A.
a2680443-664e-46d0-b4dd-97456ba810db
Ganesan, A.
62aa5a87-9308-4383-8686-99726b6bcfb9
Packham, Graham
fdabe56f-2c58-469c-aadf-38878f233394

Sharp, Adam, Crabb, Simon J., Johnson, Peter W.M., Hague, Angela, Cutress, Ramsey, Townsend, Paul A., Ganesan, A. and Packham, Graham (2009) Thioflavin S (NSC71948) interferes with Bcl-2-associated athanogene (BAG-1)-mediated protein-protein interactions. The Journal of Pharmacology and Experimental Therapeutics, 331 (2), 680-689. (doi:10.1124/jpet.109.153601).

Record type: Article

Abstract

The C-terminal BAG domain is thought to play a key role in BAG-1-induced survival and proliferation by mediating protein-protein interactions, for example, with heat shock proteins HSC70 and HSP70, and with RAF-1 kinase. Here, we have identified thioflavin S (NSC71948) as a potential small-molecule chemical inhibitor of these interactions. NSC71948 inhibited the interaction of BAG-1 and HSC70 in vitro and decreased BAG-1:HSC70 and BAG-1:HSP70 binding in intact cells. NSC71948 also reduced binding between BAG-1 and RAF-1, but had no effect on the interaction between two unrelated proteins, BIM and MCL-1. NSC71948 functionally reversed the ability of BAG-1 to promote vitamin D3 receptor-mediated transactivation, an activity of BAG-1 that depends on HSC70/HSP70 binding, and reduced phosphorylation of p44/42 mitogen-activate protein kinase. NSC71948 can be used to stain amyloid fibrils; however, structurally related compounds, thioflavin T and BTA-1, had no effect on BAG-1:HSC70 binding, suggesting that structural features important for amyloid fibril binding and inhibition of BAG-1:HSC70 binding may be separable. We demonstrated that NSC71948 inhibited the growth of BAG-1 expressing human ZR-75-1 breast cancer cells and wild-type, but not BAG-1-deficient, mouse embryo fibroblasts. Taken together, these data suggest that NSC71948 may be a useful molecule to investigate the functional significance of BAG-1 C-terminal protein interactions. However, it is important to recognize that NSC71948 may exert additional “off-target” effects. Inhibition of BAG-1 function may be an attractive strategy to inhibit the growth of BAG-1-overexpressing cancers, and further screens of additional compound collections may be warranted

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Published date: November 2009

Identifiers

Local EPrints ID: 69641
URI: http://eprints.soton.ac.uk/id/eprint/69641
ISSN: 0022-3565
PURE UUID: f7c22037-ff75-4945-8140-989e832f5f20
ORCID for Peter W.M. Johnson: ORCID iD orcid.org/0000-0003-2306-4974
ORCID for Graham Packham: ORCID iD orcid.org/0000-0002-9232-5691

Catalogue record

Date deposited: 25 Nov 2009
Last modified: 20 Jul 2019 01:12

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