!** After analyzing the single domain structure !** Modifications have been made to the following:T115, G116,L117,I119,K127,I126,L139. !** K152 has been removed as unreliable. NOEs inconsistent with structure are commented !** out but added as notes. !T112 assign (resid 112 and name hg2#) (resid 204 and name h5#) 4.0 2.2 2.5 ! T115 !*! = removed owing to uncertainty on 6/18 !*! replaced after completing structure of second domain assign (resid 115 and name hg2#) (resid 204 and name h5#) 4.0 2.2 1.5 ! weak assign (resid 115 and name hg2#) (resid 205 and name h2) 4.0 2.2 1.5 ! very weak assign (resid 115 and name og1) (resid 205 and name n6) 3.0 0.7 0.5 ! both confirmed in single domain ! G116 assign (resid 116 and name HA#) (resid 204 and name h1') 4.0 2.2 1.0 ! weak !assign (resid 116 and name HA#) (resid 204 and (name h5' or name h5'')) 4.0 2.2 1.0 ! medium assign (resid 116 and name HA#) (resid 204 and (name h2'' or name h2')) 4.0 2.2 1.0 ! weak assign (resid 116 and name HA#) (resid 204 and name h5#) 4.0 2.2 2.5 ! very weak assign (resid 116 and name ha#) (resid 205 and name h8) 4.0 2.2 2.0 assign (resid 116 and name ha#) (resid 205 and name h2) 5.0 0.5 100.0 !all of these are from the single domain complex ! L117 assign (resid 117 and name hd*) (resid 204 and name h1') 4.0 2.2 2.5 ! very weak assign (resid 117 and name hd*) (resid 204 and name h6) 4.0 2.2 2.5 ! very weak assign (resid 117 and name hd*) (resid 204 and name h5#) 4.0 2.2 3.5 ! very weak ! all in the single domain construct ! I118 ! I119 assign (resid 119 and name HD1# ) (resid 205 and name H2 ) 4.0 2.2 2.5 ! very weak ** assign (resid 119 and name HD1# ) (resid 205 and name h1') 4.0 2.2 1.5 ! medium in single domain** assign (resid 119 and name HD1# ) (resid 206 and name H5# ) 3.0 1.2 0.8 !strong in single ** assign (resid 119 and name HD1# ) (resid 206 and name h4') 4.0 2.2 2.5 ! very weak ** assign (resid 119 and name HD1# ) (resid 206 and (name H2' or name h2'')) 4.0 2.2 1.5 !strong in single assign (resid 119 and name HD1# ) (resid 207 and name H5# ) 3.0 1.2 1.3 !strong in single ** assign (resid 119 and name HD1# ) (resid 207 and name H1' ) 4.0 2.2 1.5 ! weak * assign (resid 119 and name HD1# ) ((resid 207 or resid 208) and (name h5' or name h5'')) 4.0 2.2 2.5 !weak # assign (resid 119 and name HG2# ) (resid 205 and name H1' ) 4.0 2.2 2.0 ! weak assign (resid 119 and name HG2# ) (resid 206 and name H5# ) 3.0 1.2 1.8 !medium in single ** assign (resid 119 and name HG2# ) (resid 206 and name h6) 4.0 2.2 2.5 ! weak assign (resid 119 and name HG2# ) (resid 206 and (name h2' or name h2'')) 4.0 2.2 1.5 ! strong in the single domain ** assign (resid 119 and name HG2# ) (resid 206 and name h4') 4.0 2.2 1.5 ! strong in the single domain ** assign (resid 119 and name HG2# ) (resid 207 and name H5#) 3.0 1.2 1.8 !strong in single ** assign (resid 119 and name HG2# ) (resid 207 and name h6) 4.0 2.2 2.5 ! weak assign (resid 119 and name HG2# ) (resid 207 and name H1' ) 4.0 2.2 1.5 assign (resid 119 and name HG2# ) (resid 207 and name h4') 4.0 2.2 1.5 assign (resid 119 and name HG2# ) (resid 207 and (name h2' or name h2'')) 4.0 2.2 2.5 ! very weak assign (resid 119 and name hg2#) (resid 207 and (name h5' or name h5'')) 4.0 2.2 1.5 ! strong in single cmplx * ! K121 !assign (resid 121 and name HN ) (resid 206 and (name H5' or name H5'')) 4.0 2.2 2.0 ! weak !********************** ! I commented out the stuff to the HG# as this is too overlapped with the HG's of other ! lysine residues. We can't trust it until we have a better model. The HN is going to 208 ! h5' and could also be seeing 207 h5', which I put in, but may be a stretch. ! G122 assign (resid 122 and name hn) (resid 207 and (name h5' or name h5'')) 4.0 2.2 2.0 !208 assign (resid 122 and name ha#) (resid 207 and name h3') 4.0 2.2 1.0 !*!assign (resid 122 and name hn) (resid 206 and (name h5' or name h5'')) 3.0 1.2 0.5 !4.0 2.2 2.0 ! medium !G122 hn is only going to one peak, and it is definitely C8 h5'|h5'' (as this is !resolved from C10 h5'|h5'', even though their h8's are overlapped) !I commented out the ha# because this could be to C8, C9, T7, or C12 H3' !so it would be better to have a sturucture first. the 3' are too overlapped. !single peaks assumed o.k. so I put in constraint to 208 on 7/12 ! G123 !*!assign (resid 123 and name HN ) (resid 206 and (name H5' or name H5'')) 2.5 0.7 0.2 !4.0 2.2 1.0!strong assign (resid 123 and name hn) (resid 207 and (name h5' or name h5'')) 4.0 2.2 2.0 !208 assign (resid 123 and name hn) (resid 207 and name h4') 4.0 2.2 1.0 ! E124 assign (resid 124 and name hn) (resid 207 and name h4') 4.0 2.2 2.0!weak assign (resid 124 and name hn) (resid 207 and (name h5' or name h5'')) 4.0 2.2 2.0 !weak (could be to 206) !208 ! I126 !assign (resid 126 and name hg2#) (resid 206 and name h6) 4.0 2.2 2.5 ! very weak !assign (resid 126 and name hg2#) ((resid 206 or resid 207) and name h1') 3.0 1.2 1.5 ! medium !assign (resid 126 and name hg2#) (resid 206 and name h5#) 4.0 2.2 2.0 ! weak !assign (resid 126 and name hg2#) (resid 206 and name h2') 4.0 2.2 1.5 ! weak assign (resid 126 and name hg2#) (resid 207 and name h1') 3.0 1.2 1.5 assign (resid 126 and name hg2#) (resid 207 and name h6) 4.0 2.2 2.5 ! very weak assign (resid 126 and name hg2#) (resid 207 and name h5#) 4.0 2.2 2.0 ! medium assign (resid 126 and name hg2#) ((resid 208 or resid 209) and (name h5' or name h5'')) 4.0 2.2 1.5 ! medium assign (resid 126 and name hg2#) ((resid 208 or resid 209) and name h4') 4.0 2.2 1.5 ! weak assign (resid 126 and name hg2#) ((resid 208 or resid 209) and name h1') 4.0 2.2 1.5 ! weak !all noe's from the single domain sample ! K127 assign (resid 127 and name HG# ) (resid 207 and name H1' ) 4.0 2.2 1.0 ! weak assign (resid 127 and name HG# ) ((resid 208 or resid 209) and name H1' ) 4.0 2.2 1.0 ! medium ** !assign (resid 127 and name HG# ) ((resid 208 or resid 209) and name H5 ) 4.0 2.2 2.0 ! very weak assign (resid 127 and name hg# ) ((resid 208 or resid 209) and (name h5' or name h5'')) 4.0 2.2 1.0 ! weak ** assign (resid 127 and name HG# ) ((resid 208 or resid 209) and name h4') 4.0 2.2 2.0 ! very weak assign (resid 127 and name hd#) (resid 207 and name h1') 4.0 2.2 1.0 !weak assign (resid 127 and name HD# ) ((resid 208 or resid 209) and name H4') 4.0 2.2 1.0 ! assign (resid 127 and name HD# ) ((resid 208 or resid 209 ) and (name H5' or name H5'' )) 4.0 2.2 1.0 ! medium assign (resid 127 and name hd#) ((resid 208 or resid 209) and name h1') 4.0 2.2 1.0 !medium ** assign (resid 127 and name HE# ) (resid 207 and name h1') 4.0 2.2 1.0 !weak assign (resid 127 and name HE# ) ((resid 208 or resid 209) and name H4' ) 4.0 2.2 1.0 ! strong assign (resid 127 and name HE# ) ((resid 208 or resid 209 ) and (name H5' or name H5'' )) 4.0 2.2 1.0 !4.0 2.2 1.0 ! medium ! S130 assign (resid 130 and name HB# ) (( resid 209 or resid 210) and (name H5' or name H5'')) 4.0 2.2 1.0 !4.0 2.2 2.0 !medium assign (resid 130 and name HB# ) (( resid 209 ) and name H4') 4.0 2.2 1.0 !weak assign (resid 130 and name HB# ) (( resid 209) and name h1') 4.0 2.2 2.0 !very-weak !S130 HB# could be seeing the same sequence as E124hn, or it could be seeing T11's h4' and h5'|h5'' !I choose the latter as this makes more sense. The intermol to the H1' could be either to 209 or 211 !and I choose 211 - which it actually overlays better with than 209 - because of spacing. ! R136 assign (resid 136 and name HD# ) ((resid 210) and (name h5' or name h5'')) 3.0 1.2 0.8 !3.0 1.2 0.3 !strongish assign (resid 136 and name HD# ) ((resid 209 or resid 210) and name h4') 4.0 2.2 1.0 !strongish assign (resid 136 and name HD# ) ((resid 209 ) and name H1' ) 4.0 2.2 1.0 ! medium assign (resid 136 and name hg# ) (( resid 210) and (name h5' or name h5'')) 4.0 2.2 1.0 ! medium assign (resid 136 and name HG# ) ((resid 209 or resid 210) and name h4') 4.0 2.2 1.0 !medium assign (resid 136 and name HG# ) ((resid 209 ) and name h1') 4.0 2.2 1.0 !medium assign (resid 136 and name HB# ) ((resid 209 ) and name h1') 4.0 2.2 1.0 ! L139 assign (resid 139 and name hd*) (resid 205 and name h2) 3.0 1.2 1.8 !strong in single domain hd1 assign (resid 139 and name hd*) (resid 206 and name h5#) 3.0 1.2 1.8 !strong in single domain hd1 assign (resid 139 and name hd*) (resid 207 and name h5#) 3.0 1.2 1.8 !strong in single domain hd1 !assign (resid 139 and name hd2#) (resid 205 and name h2) 4.0 2.2 2.5 ! medium !assign (resid 139 and name hd*) (resid 206 and name h5#) 4.0 2.2 1.0 !4.0 2.2 1.0 !medium !assign (resid 139 and name hd*) (resid 207 and name h5#) 4.0 2.2 2.0 ! medium-strong !****************** ! I don't see the peaks to 205 h4', h5' or h5'' under the baseline, I do see something ! that you could argue is to h5', so I left it in. There is also the same peak that I119 is ! seeing to what I am calling the h5# of 207. It is in the meth labelled sample, so I think ! that it is the real thing. assign (resid 139 and name hn) (resid 207 and name h5#) 4.0 2.2 1.5 ! K152 !assign (resid 152 and name he#) (resid 204 and name h5#) 3.0 1.2 0.3 !4.0 2.2 1.0 !medium but near a T2 ridge !assign (resid 152 and name he#) (resid 205 and name h2) 4.0 2.2 2.0 !very weak !commented this out in the single domain