Speaker
Description
Programmed cell death (apoptosis) is essential for human life. In its intrinsic pathway, the Bcl-2 (B-cell lymphoma 2) protein family regulates cell life and death by controlling permeability of the mitochondrial outer membrane (MOM). However, the molecular basis of cell protection by its anti-apoptotic Bcl-2 members remains elusive due to the lack of detailed structural insight into their action at the MOM to ensure its integrity. To provide atomic-level insight into their functioning, we will use the founding member of this family, the human antiapoptotic Bcl-2 protein itself, whose involvment in p53 regulation and its overexpression
plays a notorious role in many cancers and their treatment resistance. However, for a long time, obtaining sufficient protein was cumbersome due to its insolubility as a membrane
protein and low yields. Therefore, we establishes an expression and purification protocol [1] to produce routinely mg amounts of the fully functional full-length human isoform 2 of Bcl-2
(Bcl-2(2)) which can also be isotopically labelled as 15N/13C/2H versions, ideally suited for neutron and NMR studies. The protocol even allows to generate residue specific Bcl-2
mutants and various constructs [2].
- Aden, A.U. Mushtaq, A. Dingeldein, M. Wallgren, G. Gröbner. A novel recombinant expression and purification approach for the full-length anti-apoptotic membrane protein Bcl-2. Protein expression and purification. 172 (2020) 105628.
- A. Ul Mushtaq, J. Aden, T. Sparrman, M. Hedenstrom, G. Gröbner. Insight into Functional Membrane Proteins by Solution NMR: The Human Bcl-2 Protein-A Promising Cancer Drug Target. Molecules 26 (2021) 1467, 1-14
