Conversion of Bfl-1, an anti-apoptotic Bcl-2 family protein, to a potent pro-apoptotic protein by fusion with green fluorescent protein (GFP).
Human Bfl-1 is an anti-apoptotic Bcl-2 family member. Here, we found that Bfl-1 was converted into a potent death-promoting protein by green fluorescent protein (GFP) fusion with its N-terminus. The transient expression of GFP-Bfl-1 induced cytochrome c release and triggered apoptosis in 293T cells, which depended on the mitochondrial localization of GFP-Bfl-1. Apoptosis induced by GFP-Bfl-1 was significantly blocked by the pan-caspase inhibitor carbobenzoxy-Val-Ala-Asp-fluoromethyl ketone, but was not blocked by either Bcl-xL or Bfl-1. Our findings provide a useful model for understanding the structural basis of Bcl-2 family proteins that act in an opposite way despite sharing structural similarity between anti-apoptotic and pro-apoptotic proteins.
Ko
Jae-Kyun
JK
Cancer Research Institute, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, 110-799 Seoul, South Korea.
Choi
Kyoung-Han
KH
Kim
Hee-Jung
HJ
Choi
Hye-Young
HY
Yeo
Dong-Jun
DJ
Park
Sue-O
SO
Yang
Wan Seok
WS
Kim
Yong-Nyun
YN
Kim
Chul-Woo
CW
D017209:Apoptosis; D020169:Caspases / Q000502:physiology; D002460:Cell Line; D003574:Cytochrome c Group / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D015778:Minor Histocompatibility Antigens; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism; D051020:bcl-X Protein
C096816:BCL2-related protein A1; C494811:BCL2L1 protein, human; D003574:Cytochrome c Group; D008164:Luminescent Proteins; D015778:Minor Histocompatibility Antigens; D019253:Proto-Oncogene Proteins c-bcl-2; D011993:Recombinant Fusion Proteins; D051020:bcl-X Protein; D049452:Green Fluorescent Proteins; D020169:Caspases