U2lV[5lllTMIS43rt/| ™HÜ0O3nMli.l2
pH
SU» i? a
SUSI m
kDa 130 I—
79 k
46 h
33 h
29 h
18h
Proteomics
E 115
Q_ Q.
10
120
125
p
T»
* ^54(352 W23
S15-J* í
c.« S33C--
l^S N29 • CM*
M40     • «s
N3C
e
LM        --• ^   ... WB3 D5 " *1 EM *
136
6 7
q-r-
10
11
7($27 60^) 20°O
Q 31
25
10 O
32 64
18
14 O
56
"0 %35
29 O
3Ô2
0 O 19 ?B
16 O
Hp
52   7058 4139
oO 77 Q 60 57
36 O
4(P8
3CD2 48(3 730§65
512854 ^
O50
44j
o
5 71
O 38
k 42 53
0 0°*
82
83
78
40     *     ' *»
gT?43 'SO K24
■02,D41      f-*V3l    • •
_1_L_J
102 10.0
8.5    8.0    7.5 7.0
1H (ppm)
o o
30 37
O 40
61
76
072 179
81
Western blotting with anti-HA antibody
Input of LexA-HA proteins    Pull-down with GST-KIX
SDS-PAGE
Purified GST-KIX protein
pH
Two-dimensional gel electrophoresis
Orr
UJ	
O	_        at.* «
<	
Q. i	
Vi	
SD	
®
kDa 130
79 -
46 h
33 h
29 h
18h
15h
ISOELECTRIC FOCUSING
PH = 1ft
MASS SPECTROMETRY
5        6 7
-TTT—
10
11
7($27
eo^) 20°O
26 17 O022 1
18
1
3<&2
480 73 I 47
I o
1$?««
51 O50
44j
65
66
52   7058 4139
oO 77 O 60 57
O O 30 37
?o
O
5 71
80
6^°§® 0°
82
O 38
83
0
78
61
76
072 J079
81
Matrix-assisted laser desorption/ionization (MALDI)
HPLC High Performance Liquid Chromatography
HPLC Column
Sample Manager Solvent Manager
NMR Nuclear magnetic resonance
Radio frequency transmitter
Sweep Coils
Magnet Pole
É
Radio frequency receiver & amplifier
Sweep Coils
spinning sample tube
Magnet Pole
Control Console and Recorder
Isotope	Mag Moment (u, nm)	Nuclear Spin	Natural Abundance (%)	Gyromagnetic Ratio	NMR Frequency at 500 MHz
1H	2.79284734(3)	1/2	-100	267.522	-500
2H	0.857438228(9)	1	0.015	41.066	-76.753
3H	2.97896244(4)	1/2	0	285.349	-533.32
10B	1.80064478(6)	3	19.9	28.747	-53.718
11B	2.6886489	3/2	80.1	85.847	-160.42
13C	0.7024118(14)	1/2	1.1	67.238	-125.725
14N	0.40376100(6)	1	99.6	19.338	-36.132
15N	-0.28318884(5)	1/2	0.37	-27.126	50.782
170	-1.89379(9)	5/2	0.04	-36.281	67.782
19F	2.628868(8)	1/2	-100	251.815	-470.47
31 p	1.13160(3)	1/2	-100	108.394	-202.606
MLL
110-
CL
w
120-
"&T614 .
' A610
75*
A632» X«' T"
T"
130-
9.5
9.0
8.5
8.0
7.5
7.0
105-
110-
F6W
0
\T614
r
I. 115-1
120-
125-
130-
1* «ffl>T
A632
4
620 •A610
9.5 9.0 8.5 8.0 7.5
0)2-1H (ppm)
7.0
6.5
6.5
Chemical shift conditioned by binding
nuclear magnetic resonance MMR spectra are recorded in solution 1H-15N HSQC spectra
116
117
118
119
120
121
- /
8 8
R671
R668
F612 R624
S670 L664
pSer31
pSer32
Glu18
M639^ L652/S
E648
8.7 'H ppm
8 6
Mapping the interactions of the p53 transactivation domain with the KIX domain of CBP. Lee CW1, Arai M, Martinez-Yamout MA, Dyson HJ, Wright PE.
E2A- KIX complex
MLL - KIX complex
Leu-L
mll
m
e2a|| i
mll jj^ j
Leu
9aaTAD
mll
MLL helix
e2a
E2A helix
Gal 4
p53-tadii
p53-II helix
p53-tadi
p53-I helix
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