Quantifying the inhibitory effect of Bcl‐xl on the action of Mff using live‐cell fluorescence imaging

Autor: Yunyun Ma, Mengyan Du, Fangfang Yang, Zihao Mai, Chenshuang Zhang, Wenfeng Qu, Bin Wang, Xiaoping Wang, Tongsheng Chen
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: FEBS Open Bio, Vol 9, Iss 12, Pp 2041-2051 (2019)
Druh dokumentu: article
ISSN: 2211-5463
DOI: 10.1002/2211-5463.12739
Popis: Mitochondrial fission regulates mitochondrial function and morphology, and has been linked to apoptosis. The mitochondrial fission factor (Mff), a tail‐anchored membrane protein, induces excessive mitochondrial fission, contributing to mitochondrial dysfunction and apoptosis. Here, we evaluated the inhibitory effect of Bcl‐xl, an antiapoptotic protein, on the action of Mff by using live‐cell fluorescence imaging. Microscopic imaging analysis showed that overexpression of Mff induced mitochondrial fragmentation and apoptosis, which were reversed by coexpression of Bcl‐xl. Microscopic imaging and live‐cell fluorescence resonance energy transfer analysis demonstrated that Bcl‐xl reconstructs the Mff network from punctate distribution of higher‐order oligomers to filamentous distribution of lower‐order oligomers. Live‐cell fluorescence resonance energy transfer two‐hybrid assay showed that Bcl‐xl interacted with Mff to form heterogenous oligomers with 1 : 2 stoichiometry in cytoplasm and 1 : 1 stoichiometry on mitochondria, indicating that two Bcl‐xl molecules primarily interact with four Mff molecules in cytoplasm, but with two Mff molecules on mitochondria.
Databáze: Directory of Open Access Journals