TY - JOUR
T1 - Reconstitution and functional characterization of the FtsH protease in lipid nanodiscs
AU - Prabudiansyah, Irfan
AU - van der Valk, Ramon
AU - Aubin-Tam, Marie Eve
PY - 2021
Y1 - 2021
N2 - FtsH is a membrane-bound protease that plays a crucial role in proteolytic regulation of many cellular functions. It is universally conserved in bacteria and responsible for the degradation of misfolded or misassembled proteins. A recent study has determined the structure of bacterial FtsH in detergent micelles. To properly study the function of FtsH in a native-like environment, we reconstituted the FtsH complex into lipid nanodiscs. We found that FtsH in membrane scaffold protein (MSP) nanodiscs maintains its native hexameric conformation and is functionally active. We further investigated the effect of the lipid bilayer composition (acyl chain length, saturation, head group charge and size) on FtsH proteolytic activity. We found that the lipid acyl chain length influences AaFtsH activity in nanodiscs, with the greatest activity in a bilayer of di-C18:1 PC. We conclude that MSP nanodiscs are suitable model membranes for further in vitro studies of the FtsH protease complex.
AB - FtsH is a membrane-bound protease that plays a crucial role in proteolytic regulation of many cellular functions. It is universally conserved in bacteria and responsible for the degradation of misfolded or misassembled proteins. A recent study has determined the structure of bacterial FtsH in detergent micelles. To properly study the function of FtsH in a native-like environment, we reconstituted the FtsH complex into lipid nanodiscs. We found that FtsH in membrane scaffold protein (MSP) nanodiscs maintains its native hexameric conformation and is functionally active. We further investigated the effect of the lipid bilayer composition (acyl chain length, saturation, head group charge and size) on FtsH proteolytic activity. We found that the lipid acyl chain length influences AaFtsH activity in nanodiscs, with the greatest activity in a bilayer of di-C18:1 PC. We conclude that MSP nanodiscs are suitable model membranes for further in vitro studies of the FtsH protease complex.
KW - AAA+ protease
KW - Lipid bilayer
KW - Membrane protein
KW - Nanodisc
KW - Proteolysis
UR - http://www.scopus.com/inward/record.url?scp=85097471455&partnerID=8YFLogxK
U2 - 10.1016/j.bbamem.2020.183526
DO - 10.1016/j.bbamem.2020.183526
M3 - Article
C2 - 33278347
AN - SCOPUS:85097471455
SN - 0005-2736
VL - 1863
JO - Biochimica et Biophysica Acta - Biomembranes
JF - Biochimica et Biophysica Acta - Biomembranes
IS - 2
M1 - 183526
ER -