High yield purification of an Isoleucine zipper modified CD95 Ligand with either biotin or DNA-oligomer binding domain for efficient Cell Apoptosis Induction DOI Creative Commons

Xiaoyue Shang,

Nina Bartels, Johann M. Weck

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 13, 2024

Abstract Background Cluster of differentiation 95 (CD95/Fas/Apo1) as part the Tumor-necrosis factor (TNF) receptor family is a prototypic trigger ‘extrinsic’ apoptotic pathway and its activation by trimeric ligand CD95L high interest for anticancer therapy. However, CD95L, when presented in solution, exhibits low efficiency to induce apoptosis signaling human cells. Results Here, we design recombinant exhibiting an isoleucine zipper (IZ) motif at N-terminus stabilization trimerized demonstrate induction efficiency. A cysteine amino acid fused behind IZ further used versatile coupling site bionanotechnological applications or development biomedical assays. fast, cheap, high-yield production via HEK293T secretory expression system presented, along with affinity purification functionalization. We verified biological activity purified protein identified stabilized structure most potent inducer signaling. Conclusions The workflow findings reported here will streamline wide array future low- high-throughput TNF-ligand screens, their modification towards improving

Language: Английский

High yield purification of an Isoleucine zipper modified CD95 Ligand with either biotin or DNA-oligomer binding domain for efficient Cell Apoptosis Induction DOI Creative Commons

Xiaoyue Shang,

Nina Bartels, Johann M. Weck

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 13, 2024

Abstract Background Cluster of differentiation 95 (CD95/Fas/Apo1) as part the Tumor-necrosis factor (TNF) receptor family is a prototypic trigger ‘extrinsic’ apoptotic pathway and its activation by trimeric ligand CD95L high interest for anticancer therapy. However, CD95L, when presented in solution, exhibits low efficiency to induce apoptosis signaling human cells. Results Here, we design recombinant exhibiting an isoleucine zipper (IZ) motif at N-terminus stabilization trimerized demonstrate induction efficiency. A cysteine amino acid fused behind IZ further used versatile coupling site bionanotechnological applications or development biomedical assays. fast, cheap, high-yield production via HEK293T secretory expression system presented, along with affinity purification functionalization. We verified biological activity purified protein identified stabilized structure most potent inducer signaling. Conclusions The workflow findings reported here will streamline wide array future low- high-throughput TNF-ligand screens, their modification towards improving

Language: Английский

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