http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Ganglioside-Embedded Nanodisc Inhibits Influenza A Virus Infection by Perforating Viral Envelope
Byoungjae KONG,Yuna KIM,Seokoh MOON,Younghun JUNG,Jonghyeok SHIN,Joon-bum PARK,Myungseo PARK,Jiwon YU,Seok-Hyeon YU,Chakhee KIM,Hooyeon KIM,Sora CHO,Yun Jeong PARK,Dae-Hyuk KWEON 한국생물공학회 2017 한국생물공학회 학술대회 Vol.2017 No.10
Perforation of Membrane Envelope of Influenza A Virus by a Ganglioside-embedded Nanoparticle
Seokoh MOON,Byoungjae KONG,Yuna KIM,Younghun JUNG,Seok-Hyeon YU,Choongjin BAN,Jonghyeok SHIN,Joonbum PARK,Myungseo PARK,Chakhee KIM,Hooyeon KIM,Sora CHO,Yunjeong PARK,Hye Rin KIM,Hyun Seok OH,Jin Kyeo 한국생물공학회 2018 한국생물공학회 학술대회 Vol.2018 No.10
Green fluorescence protein-based content-mixing assay of SNARE-driven membrane fusion
Heo, Paul,Kong, Byoungjae,Jung, Young-Hun,Park, Joon-Bum,Shin, Jonghyeok,Park, Myungseo,Kweon, Dae-Hyuk Academic Press 2017 Biochemical and biophysical research communication Vol. No.
<P><B>Abstract</B></P> <P>Soluble <I>N</I>-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins mediate intracellular membrane fusion by forming a ternary SNARE complex. A minimalist approach utilizing proteoliposomes with reconstituted SNARE proteins yielded a wealth of information pinpointing the molecular mechanism of SNARE-mediated fusion and its regulation by accessory proteins. Two important attributes of a membrane fusion are lipid-mixing and the formation of an aqueous passage between apposing membranes. These two attributes are typically observed by using various fluorescent dyes. Currently available <I>in vitro</I> assay systems for observing fusion pore opening have several weaknesses such as cargo-bleeding, incomplete removal of unencapsulated dyes, and inadequate information regarding the size of the fusion pore, limiting measurements of the final stage of membrane fusion. In the present study, we used a biotinylated green fluorescence protein and streptavidin conjugated with Dylight 594 (DyStrp) as a Föster resonance energy transfer (FRET) donor and acceptor, respectively. This FRET pair encapsulated in each v-vesicle containing synaptobrevin and t-vesicle containing a binary acceptor complex of syntaxin 1a and synaptosomal-associated protein 25 revealed the opening of a large fusion pore of more than 5 nm, without the unwanted signals from unencapsulated dyes or leakage. This system enabled determination of the stoichiometry of the merging vesicles because the FRET efficiency of the FRET pair depended on the molar ratio between dyes. Here, we report a robust and informative assay for SNARE-mediated fusion pore opening.</P> <P><B>Highlights</B></P> <P> <UL> <LI> SNARE proteins drive membrane fusion and open a pore for cargo release. </LI> <LI> Biotinylated GFP and DyStrp was used as the reporter pair of fusion pore opening. </LI> <LI> Procedure for efficient SNARE reconstitution and reporter encapsulation was established. </LI> <LI> The FRET pair reported opening of a large fusion pore bigger than 5 nm. </LI> <LI> The assay was robust and provided information of stoichiometry of vesicle fusion. </LI> </UL> </P>