LECTURE 8 TOCSY-HSQC i 4>2 1 j_ y 1lj | ti ||DIPSY-2rc| 4J | 4J | 1 1 4J 1 I t2 5N =x,x5-x5-x5x5x,-x5-x (t)3=x5-x5x5-x5x5-x5x,-x ^i=y.y-y-y.y.y-y-y fe=y-y.y-y.y-y.y-y (|)2=x5x,x,x,-x,-x5-x5-x receiver phase: x,-x,-x,x,-x5x,x5-x 4J 1 t3 Ik I1 1 GARP HC(C)H-TOCSY 4>i/4>H 1V 1 H |4J ti |4J 1 3C x 1 3C0 t2 t2 DIPSY-2 <|>1 =X,-X,X,-X,X,-X,X,-X (|)2=X,X,-X,-X,X,X,-X,-X 4>i =y-y.y-y.y-y.y-y 4>2=y.y -y -y.y.y -y -y receiver phase: x,-x,-x,x,-x,x,x,-x 1 1» |4J t3 ' /V VVV 'V GARP NOESY-HSQC i §2 lultl 1 1 y m 1 J 4J I 4J I I 4J 5N =x,x,-x5-x,x5x5-x,-x (|>3=x,-x,x,-x,x,-x,x,-x 4>i=y,y-y-y,y,y-y-y y-y>y-y>y-y (t)2=x,x5x5x5-x5-x5-x5-x receiver phase: x,-x5-x5x5-x,x5x5-x 9.0 8.5 8.0 7.5 7.0 6.5 J_I_I_L__I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_l_ tt 82 (15N) : 113.721 ppm o o i i i i i i i i i 8 0 I I I I I I I I I I I 8.0 7.5 7.0 i i i i h0.5 H.o H.5 h2.0 h2.5 h3.0 9.0 8.5 83 (1H)/ppm 6.5 geminal protons in methylene H-C-H 0.17 nm vicinal protons in aromatic ring H-C=C-H 0.25 nm Hf and H^J_3 protons in a-helix 0.34 nm meta protons in aromatic ring H-C=CH-C-H 0.42 nm Data /3-sheet a-helix ô (CO) <5(Ha) t t I l_iai_i N ljNljN Hi HH-1 0.22 nm 0.40 nm 0.35 nm 0.28 nm l_iq|_| l\ l_iq|_| l\ l_iq|_| l\ -2 -3I -4 too far too far too far 0.42 nm 0.34 nm 0.42 nm 3J(hNh?) > 8 Hz < 5 Hz -180 -120 -60 0 60 120 180 / deg. TALOS prediction.......(oo) . . . . (ooooooooo) . . . <=>. (oooooooo) , , , , (oo oooooooooooo)............................ 3J prediction .......(ooo).......(oooo)..........(oooooooo........ooooooooo).......=>. . <=............... NOE prediction ........(oooo) . . (oooooooooooooo.ooooooooooooo) . . (o oooooooooooooo)..............(oo) . < >. CSI prediction .......(ooo) . . . (ooooooooooo) . . . . (ooooooooooo) . . (oo oooooooooooooooo)........................ NN(i,i + 1 aN(i,i + 1 aN(i,i + 2) aN(i,i + 3) aN(i,i + 4) 10 20 30 40 50 60 70 80 90 GIKQYSQEELKEMALVEIAHELFEEHKKPVPFQELLNEIASLLGVKKEEL GDRIAQFYTDLNIDGRFLALSDQTWGLRSWYPYDQLDEETQLEHHHHHH TALOS -----«0000 — ««00000000000---•••-oooooooooo — ••ooo -oooooo-ooooo-----•------•------•----------------- 3J(HaH^) o o - • - o o o o o • - o o o • - o o o o o o — o« • 00*000000000 ioooi - o - o o o o o o o o - • o -•-••oom ooo*---i-oo-o---• - CO CSI ••—ooooooo—oo«oooooooooo« - -ooooooo-ooo-o«oooo oo-o-o—ooo-o- oo**-o-**** -oo- —••-•oo—o-«o o Ca CSI tto-nooooot — ooooooooooooo»-»-ooooooooooo-o«iooo -oooooo-ooooo»ooo#»«io-o«»noo»i-t----oo-----oo Ha CSI -#oot-ooooo-o-o-iooo-o#t — o»»noo-ooooooo---t-oo- — o-ooo«oooooio-o«io-oooiio«ooo« — oo#ooooo-o«o CSI -otioon----o-«-«-«« — • — ••o-o-o«---• — • — o-o — •- —•o-««-« — tooo — ooi — ••ooo*-------*----o* — ••• consensus CSI ---o---ooooo---0-00000000000----ooooooooooo-o — ooo -oooooo-ooooo-ooo----o-o---too--------oo----• o---