MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQT

1. Nuclear magnetic resonance (NMR) (318 K; pH: 6; Syb(1-116) (13C- and 15N-labelled protein); DPC (lipid) 200 mM; NaCl 150 mM; DTT 5 mM; EDTA 1 mM; Mes 20 mM)


2. Circular dichroism (CD) spectroscopy, far-UV (277 K; pH: 7.4; DTT (Standard buffer) 1 mM; EDTA (Standard buffer) 1 mM; NaCl (Standard buffer) 100 mM; Syb (protein) 4.7 uM; Tris (Standard buffer) 20 mM)


3. Circular dichroism (CD) spectroscopy, far-UV (298 K; pH: 7; 2-mercaptoethanol (buffer) 1 mM; β-octyl glucoside (detergent) 50 mM; NaCl (buffer) 50 mM; sodium phosphate (buffer) 10 mM; Syb (protein) 100 uM)


4. Fourier transform infrared spectroscopy (FTIR), aka infrared spectroscopy) (298 K; pH: 7; β-octyl glucoside (detergent) 5 %; POPC liposomes (1:100 protein-to-lipid ratio); Syb protein)

1. Ellena JF, Liang B, Wiktor M, Stein A, Cafiso DS, Jahn R, Tamm LK. "Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation." Proc Natl Acad Sci U S A. 2009; 106(48): 20306-11. PubMed: 19918058
   
   
2. Fasshauer D, Otto H, Eliason WK, Jahn R, Brunger AT. "Structural changes are associated with soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex formation." J Biol Chem. 1997; 272(44): 28036-41. PubMed: 9346956
   
   

Additional reference: Bowen M, et al. (2006) PMID 16709671. Additional reference: Stein A, et al (2009) PMID 19571812.

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Sources of Detection Methods are as follows: 1. NMR: Ellena et al (2009); 2. CD at 277K: Fasshauer et al (1997); 3. CD at 298K: Bowen et al (2006); 4. FTIR: Bowen et al (2006)

QAQVDEVVDIMRVNVDKVL

1. Ellena JF, Liang B, Wiktor M, Stein A, Cafiso DS, Jahn R, Tamm LK. "Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation." Proc Natl Acad Sci U S A. 2009; 106(48): 20306-11. PubMed: 19918058
   
   

See Region 1 for Detection Methods, Detection Method sources and additional reference information.

ERDQKLSELDDRADALQAGASQ

1. Ellena JF, Liang B, Wiktor M, Stein A, Cafiso DS, Jahn R, Tamm LK. "Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation." Proc Natl Acad Sci U S A. 2009; 106(48): 20306-11. PubMed: 19918058
   
   

See Region 1 for Detection Methods, Detection Method sources and additional reference information.

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According to Ellena et al. (2009), Region 3 "may serve as a 'stop-folding' signal."

FETSAAKLKRKY

1. Ellena JF, Liang B, Wiktor M, Stein A, Cafiso DS, Jahn R, Tamm LK. "Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation." Proc Natl Acad Sci U S A. 2009; 106(48): 20306-11. PubMed: 19918058
   
   

See Region 1 for Detection Methods, Detection Method sources and additional reference information.

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According to Ellena et al. (2009), Region 4 (Helix II) "may well transmit force from SNARE complex folding into the membrane and thus coerce the two membranes to fuse..."

WWKN

1. Ellena JF, Liang B, Wiktor M, Stein A, Cafiso DS, Jahn R, Tamm LK. "Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation." Proc Natl Acad Sci U S A. 2009; 106(48): 20306-11. PubMed: 19918058
   
   

See Region 1 for Detection Methods, Detection Method sources and additional reference information.

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This region "forms a flexible hinge at the micelle surface," according to Ellena et al. (2009), and it "appears to be ideally designed to convert a 'trans'-SNARE into a 'cis'-SNARE complex".

LKMMIILGVICAIILIIIIVYFS

1. Ellena JF, Liang B, Wiktor M, Stein A, Cafiso DS, Jahn R, Tamm LK. "Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation." Proc Natl Acad Sci U S A. 2009; 106(48): 20306-11. PubMed: 19918058
   
   

This ordered helix is the transmembrane (TM) domain.

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See Region 1 for Detection Methods, Detection Method sources and additional reference information.

1. Ellena JF, Liang B, Wiktor M, Stein A, Cafiso DS, Jahn R, Tamm LK. "Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation." Proc Natl Acad Sci U S A. 2009; 106(48): 20306-11. PubMed: 19918058
   
   
2. Fasshauer D, Otto H, Eliason WK, Jahn R, Brunger AT. "Structural changes are associated with soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex formation." J Biol Chem. 1997; 272(44): 28036-41. PubMed: 9346956
   
   

Additional references: Bowen M, et al. (2006) PMID 16709671; Stein A, et al (2009) PMID 19571812.

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Additional PDB structures of the SNARE complex featuring a fragment of DP00622 (aa 30-116) are 3HDT and 3IPD, from Stein et al (2009).

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Sent for AV (9-14-2010) PubMed: 19918058