A novel recombinant expression and purification approach for the full-length anti-apoptotic membrane protein Bcl-2

Programmed cell death (apoptosis) is an essential mechanism in life that tightly regulates embryogenesis and removal of harmful cells. Besides an extrinsic pathway, an intrinsic (mitochondrial) apoptotic pathway exists where mitochondria are actively involved in cellular clearance in response to internal stress signals. Pro-apoptotic (death) and anti-apoptotic (survival) members of the B cell CLL/lymphoma-2 (Bcl-2) protein family meet at the mitochondrion's surface where they accurately regulate apoptosis. Overexpression of the anti-apoptotic Bcl-2 protein is a hallmark for many types of cancers and in particular for many treatment resistant tumors. Bcl-2 is a membrane protein residing in the mitochondrial outer membrane. Due to its typical membrane protein features including very limited solubility, it is difficult to express and to purify. Therefore, most biophysical and structural studies have used truncated, soluble versions. However, to understand its membrane-coupled function and structure, access to sufficient amount of full-length human Bcl-2 protein is a necessity. Here, we present a novel, E. coli based approach for expression and purification of preparative amounts of the full-length human isoform 2 of Bcl-2 (Bcl-2(2)), solubilized in detergent micelles, which allows for easy exchange of the detergent.

Ul Mushtaq, AmeeqUmeå universitet,Kemiska institutionen(Swepub:umu)amul0005 (författare)
Dingeldein, Artur P. G.Umeå universitet,Kemiska institutionen(Swepub:umu)ardi0002 (författare)
Wallgren, Marcus,1978-Umeå universitet,Institutionen för medicinsk kemi och biofysik(Swepub:umu)muswan02 (författare)
Gröbner, GerhardUmeå universitet,Kemiska institutionen(Swepub:umu)gegr0001 (författare)
Umeå universitetKemiska institutionen (creator_code:org_t)

Av författaren/redakt...: Ådén, Jörgen, 19 ...; Ul Mushtaq, Amee ...; Dingeldein, Artu ...; Wallgren, Marcus ...; Gröbner, Gerhard

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MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Medicinsk biotek ...; och Medicinsk biotek ...

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