Three αSNAP and 10 ATP Molecules Are Used in SNARE Complex Disassembly by N-ethylmaleimide-sensitive Factor (NSF)

Shah, Niket; Colbert, Karen N.; Enos, M.D.; Herschlag, Daniel; Weis, William I.

doi:10.1074/jbc.m114.620849

Cited by 20 publications

(20 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consequently, membranes may attenuate or inhibit association of α-SNAP to the LD, which reduces the negative role of α-SNAP in LD zippering and leads to a membrane curvature-dependent role of α-SNAP in membrane fusion (Park et al, 2014; Zick et al, 2015). Finally, our data support the finding that α-SNAP mainly binds the cytoplasmic SNARE 4HB in a 1:1 stoichiometry at up to a 10-μM α-SNAP concentration (Shah et al, 2015; Vivona et al, 2013). However, NSF and/or membranes promote up to four α-SNAP molecules to bind each SNARE 4HB (Zhao et al, 2015).…”

Section: Discussionsupporting

confidence: 88%

“…Second, α-SNAP increased the SNARE CTD zippering energy by ∼12.4 k B T and decreased the LD zippering energy by ∼8 k B T under standard conditions (calculated from ∼4 μM dissociation constant), leading to a net energy gain of ∼4.4 k B T (Gao et al, 2012). This energy increase is expected to significantly enhance membrane fusion (Ma et al, 2016), but it does not likely impede SNARE disassembly because the energy gain is negligible compared to the ∼ 70 k B T total SNARE zippering energy (Gao et al, 2012; Ma et al, 2016) and the ∼200 k B T total energy necessary for NSF to disassemble a single SNARE complex (Shah et al, 2015). Third, the dual role of α-SNAP in SNARE zippering may cause the observed positive and negative effects of α-SNAP on membrane fusion (Park et al, 2014; Stein et al, 2009; Zick et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…α-SNAP alone primarily binds cytoplasmic SNARE complexes in a 1:1 stoichiometry. However, in the presence of membranes or NSF, α-SNAP is found to associate with SNARE complexes in different stoichiometries, ranging from one to four α-SNAP molecules per SNARE complex (Marz et al, 2003; Shah et al, 2015; Wimmer et al, 2001; Zhao et al, 2015; Zhou et al, 2015). How α-SNAP binds SNARE complexes with different stoichiometries remains mysterious.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

α-SNAP Enhances SNARE Zippering by Stabilizing the SNARE Four-Helix Bundle

Kang

Jiao

et al. 2016

Cell Reports

View full text Add to dashboard Cite

Summary Intracellular membrane fusion is mediated by dynamic assembly and disassembly of soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein (SNAP) receptors (SNAREs). α-SNAP guides NSF to disassemble SNARE complexes after membrane fusion. Recent experiments showed that α-SNAP also dramatically enhances SNARE assembly and membrane fusion. How α-SNAP plays roles in these opposing activities is not known. Here, we examined the effect of α-SNAP on the step-wise assembly of the synaptic SNARE complex using optical tweezers. We found that α-SNAP destabilized the linker domain (LD) of the SNARE complex but stabilized its C-terminal domain (CTD) through a conformational selection mechanism. In contrast, α-SNAP minimally affected assembly of the SNARE N-terminal domain (NTD), indicating that α-SNAP barely bound the partially assembled trans-SNARE complex. Thus, α-SNAP recognizes the folded CTD for SNARE disassembly with NSF and subtly modulates membrane fusion by altering the stabilities of the SNARE CTD and LD.

show abstract

Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

α-SNAP Enhances SNARE Zippering by Stabilizing the SNARE Four-Helix Bundle

Kang

Jiao

et al. 2016

Cell Reports

View full text Add to dashboard Cite

show abstract

“…Some studies suggest that the disassembly of a single SNARE by NSF is a processive process with the consumption of ATP molecules ranging from 10 to 50 [33,34]. An alternative model suggests that turnover of a single ATP is sufficient to disassemble a SNARE complex, indicating a conformational switch of NSF upon ATP hydrolysis to cause the SNARE disassembly [33].…”

Section: Discussionmentioning

confidence: 99%

Cryo-EM structure of SNAP-SNARE assembly in 20S particle

et al. 2015

View full text Add to dashboard Cite

N-ethylmaleimide-sensitive factor (NSF) and α soluble NSF attachment proteins (α-SNAPs) work together within a 20S particle to disassemble and recycle the SNAP receptor (SNARE) complex after intracellular membrane fusion. To understand the disassembly mechanism of the SNARE complex by NSF and α-SNAP, we performed single-particle cryo-electron microscopy analysis of 20S particles and determined the structure of the α-SNAP-SNARE assembly portion at a resolution of 7.35 Å. The structure illustrates that four α-SNAPs wrap around the single left-handed SNARE helical bundle as a right-handed cylindrical assembly within a 20S particle. A conserved hydrophobic patch connecting helices 9 and 10 of each α-SNAP forms a chock protruding into the groove of the SNARE four-helix bundle. Biochemical studies proved that this structural element was critical for SNARE complex disassembly. Our study suggests how four α-SNAPs may coordinate with the NSF to tear the SNARE complex into individual proteins.

show abstract

“…The estimated number of ATP molecules required for SNARE complex disassembly ranges from 10 to 50 [38,39]. A recent study demonstrated that a set of initially bound ATP molecules are sufficient for complete SNARE complex disassembly, i.e.…”

Section: Mechanism Of Nsf Mediated Snare Complex Disassemblymentioning

confidence: 99%

Recent Advances in Deciphering the Structure and Molecular Mechanism of the AAA+ ATPase N-Ethylmaleimide-Sensitive Factor (NSF)

Zhao

Brunger

2016

Journal of Molecular Biology

View full text Add to dashboard Cite

NSF (N-ethylmaleimide Sensitive Factor), first discovered in 1988, is a key factor for eukaryotic trafficking, including protein and hormone secretion and neurotransmitter release. It is a member of the AAA+ family (ATPases Associated with diverse cellular Activities). NSF disassembles SNARE (Soluble N-ethylmaleimide sensitive factor Attachment protein REceptors) complexes in conjunction with SNAP (Soluble N-ethylmaleimide sensitive factor Attachment Protein) adaptor proteins. Structural studies of NSF and its complex with SNARE and SNAP (known as 20S supercomplex) started about twenty years ago. Crystal structures of individual N and D2 domains of NSF and low-resolution electron microscopy structures of full-length NSF and 20S supercomplex have been reported over the years. Nevertheless, the molecular architecture of the 20S supercomplex and the molecular mechanism of NSF-mediated SNARE complex disassembly remained unclear until recently. Here we review recent atomic-resolution or near-atomic resolution structures of NSF and of the 20S supercomplex, and recent insights into the molecular mechanism and energy requirements of NSF. We also compare NSF with other known AAA+ family members.

show abstract

Three αSNAP and 10 ATP Molecules Are Used in SNARE Complex Disassembly by N-ethylmaleimide-sensitive Factor (NSF)

Cited by 20 publications

References 33 publications

α-SNAP Enhances SNARE Zippering by Stabilizing the SNARE Four-Helix Bundle

α-SNAP Enhances SNARE Zippering by Stabilizing the SNARE Four-Helix Bundle

Cryo-EM structure of SNAP-SNARE assembly in 20S particle

Recent Advances in Deciphering the Structure and Molecular Mechanism of the AAA+ ATPase N-Ethylmaleimide-Sensitive Factor (NSF)

Contact Info

Product

Resources

About