A search for synthetic peptides that inhibit soluble N-ethylmaleimide sensitive-factor attachment receptor-mediated membrane fusion.
Soluble N-ethylmaleimide sensitive-factor attachment receptor (SNARE) proteins have crucial roles in driving exocytic membrane fusion. Molecular recognition between vesicle-associated (v)-SNARE and target membrane (t)-SNARE leads to the formation of a four-helix bundle, which facilitates the merging of two apposing membranes. Synthetic peptides patterned after the SNARE motifs are predicted to block SNARE complex formation by competing with the parental SNAREs, inhibiting neuronal exocytosis. As an initial attempt to identify the peptide sequences that block SNARE assembly and membrane fusion, we created thirteen 17-residue synthetic peptides derived from the SNARE motifs of v- and t-SNAREs. The effects of these peptides on SNARE-mediated membrane fusion were investigated using an in vitro lipid-mixing assay, in vivo neurotransmitter release and SNARE complex formation assays in PC12 cells. Peptides derived from the N-terminal region of SNARE motifs had significant inhibitory effects on neuroexocytosis, whereas middle- and C-terminal-mimicking peptides did not exhibit much inhibitory function. N-terminal mimicking peptides blocked N-terminal zippering of SNAREs, a rate-limiting step in SNARE-driven membrane fusion. Therefore, the results suggest that the N-terminal regions of SNARE motifs are excellent targets for the development of drugs to block SNARE-mediated membrane fusion and neurotransmitter release.
Jung
Chang H
CH
School of Biotechnology and Bioengineering, Sungkyunkwan University, Gyeonggi-do, Korea.
Yang
Yoo-Soo
YS
Kim
Jun-Seob
JS
Shin
Jae-Il
JI
Jin
Yong-Su
YS
Shin
Jae Y
JY
Lee
Jong H
JH
Chung
Koo M
KM
Hwang
Jae S
JS
Oh
Jung M
JM
Shin
Yeon-Kyun
YK
Kweon
Dae-Hyuk
DH
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D005089:Exocytosis / Q000187:drug effects*; D007700:Kinetics; D008561:Membrane Fusion / Q000187:drug effects; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D009638:Norepinephrine / Q000378:metabolism; D016716:PC12 Cells; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D050600:SNARE Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry
D010455:Peptides; D050600:SNARE Proteins; D009638:Norepinephrine