Dataset Information

Selective Affimers Recognise the BCL-2 Family Proteins BCL-x_L and MCL-1 through Noncanonical Structural Motifs*.

ABSTRACT: The BCL-2 family is a challenging group of proteins to target selectively due to sequence and structural homologies across the family. Selective ligands for the BCL-2 family regulators of apoptosis are useful as probes to understand cell biology and apoptotic signalling pathways, and as starting points for inhibitor design. We have used phage display to isolate Affimer reagents (non-antibody-binding proteins based on a conserved scaffold) to identify ligands for MCL-1, BCL-x_L , BCL-2, BAK and BAX, then used multiple biophysical characterisation methods to probe the interactions. We established that purified Affimers elicit selective recognition of their target BCL-2 protein. For anti-apoptotic targets BCL-x_L and MCL-1, competitive inhibition of their canonical protein-protein interactions is demonstrated. Co-crystal structures reveal an unprecedented mode of molecular recognition; where a BH3 helix is normally bound, flexible loops from the Affimer dock into the BH3 binding cleft. Moreover, the Affimers induce a change in the target proteins towards a desirable drug-bound-like conformation. These proof-of-concept studies indicate that Affimers could be used as alternative templates to inspire the design of selective BCL-2 family modulators and more generally other protein-protein interaction inhibitors.

SUBMITTER: Miles JA

PROVIDER: S-EPMC7821230 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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Publications

Selective Affimers Recognise the BCL-2 Family Proteins BCL-x<sub>L</sub> and MCL-1 through Noncanonical Structural Motifs*.

Miles Jennifer A JA Hobor Fruzsina F Trinh Chi H CH Taylor James J Tiede Christian C Rowell Philip R PR Jackson Brian R BR Nadat Fatima A FA Ramsahye Pallavi P Kyle Hannah F HF Wicky Basile I M BIM Clarke Jane J Tomlinson Darren C DC Wilson Andrew J AJ Edwards Thomas A TA

Chembiochem : a European journal of chemical biology 20201102 1

The BCL-2 family is a challenging group of proteins to target selectively due to sequence and structural homologies across the family. Selective ligands for the BCL-2 family regulators of apoptosis are useful as probes to understand cell biology and apoptotic signalling pathways, and as starting points for inhibitor design. We have used phage display to isolate Affimer reagents (non-antibody-binding proteins based on a conserved scaffold) to identify ligands for MCL-1, BCL-x<sub>L</sub> , BCL-2, ...[more]

PMID: 32961017

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Project description:Development of resistance to chemo- and radiotherapy in patients suffering from advanced cervical cancer narrows the therapeutic window for conventional therapies. Previously we reported that a combination of the selective BCL-2 family inhibitors ABT-263 and A-1210477 decreased cell proliferation in C33A, SiHa and CaSki human cervical cancer cell lines. As ABT-263 binds to both BCL-2 and BCL-XL with high affinity, it was unclear whether the synergism of the drug combination was driven either by singly inhibiting BCL-2 or BCL-XL, or inhibition of both. In this present study, we used the BCL-2 selective inhibitor ABT-199 and the BCL-XL selective inhibitor A1331852 to resolve the individual antitumor activities of ABT-263 into BCL-2 and BCL-XL dependent mechanisms. A-1210477 was substituted for the orally bioavailable S63845. Four cervical cancer cell lines were treated with the selective BCL-2 family inhibitors ABT-199, A1331852 and S63845 alone and in combination using 2-dimensional (2D) and 3-dimensional (3D) cell culture models. The SiHa, C33A and CaSki cell lines were resistant to single agent treatment of all three drugs, suggesting that none of the BCL-2 family of proteins mediate survival of the cells in isolation. HeLa cells were resistant to single agent treatment of ABT-199 and A1331852 but were sensitive to S63845 indicating that they depend on MCL-1 for survival. Co-inhibition of BCL-2 and MCL-1 with ABT-199 and S63845, inhibited cell proliferation of all cancer cell lines, except SiHa. However, the effect of the combination was not as pronounced as combination of A1331852 and S63845. Co-inhibition of BCL-XL and MCL-1 with A1331852 and S63845 significantly inhibited cell proliferation of all four cell lines. Similar data were obtained with 3-dimensional spheroid cell culture models generated from two cervical cancer cell lines in vitro. Treatment with a combination of A1331852 and S63845 resulted in inhibition of growth and invasion of the 3D spheroids. Collectively, our data demonstrate that the combination of MCL-1-selective inhibitors with either selective inhibitors of either BCL-XL or BCL-2 may be potentially useful as treatment strategies for the management of cervical cancer.

Dataset Information

Selective Affimers Recognise the BCL-2 Family Proteins BCL-x_L and MCL-1 through Noncanonical Structural Motifs*.

Publications

Selective Affimers Recognise the BCL-2 Family Proteins BCL-x<sub>L</sub> and MCL-1 through Noncanonical Structural Motifs*.

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Dataset Information

Selective Affimers Recognise the BCL-2 Family Proteins BCL-xL and MCL-1 through Noncanonical Structural Motifs*.

Publications

Selective Affimers Recognise the BCL-2 Family Proteins BCL-x<sub>L</sub> and MCL-1 through Noncanonical Structural Motifs*.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Selective Affimers Recognise the BCL-2 Family Proteins BCL-x_L and MCL-1 through Noncanonical Structural Motifs*.