4
Analysis of Intact Macromolecular Assemblies On A Bench Top Orbitrap MS System
tate to study
using a
mass
ray using an
s of various
e resp. isoform
vation even
g ESI-MS
nditions,
mblies and
conditions, fully
ut these
ited surface
t physiological
lues, which
apable of
achievable
cific isoforms or
s EMR bench
tect ion signals
power thus
d precision for
Results
Carbonic Anhydrase
Smaller proteins can easily measured by mass
spectrometry, but under native conditions even these show
quite different behavior. The signal of carbonic anhydrase
for example, as member of this group of proteins, is
reduced to two major charge states, making a classical
deconvolution based on charge envelop pattern difficult.
Due to the high resolution of the Orbitrap detection
system, this 29 kDa protein still can be isotopically
resolved (see fig. 2).
Herceptin
GroEL
The chaperone pro
group large protein
fulllyactive 14-mer
resolving power of t
minor part for succ
conjugates, but des
factor. With GroEL
resulting in sharp b
charge states (see
FIGURE 2. Spectrum of carbonic anhydrase in native
state. Isotopic resolution is easily achieved.
carbonic_140k_inf_003
#
18-26
RT:
0.54-0.78
AV:
9
NL:
9.09E4
T:
FTMS+pNSIsid=1.00 Fullms21000.00@hcd100.00 [500.00-20000.00]
2600
2700
2800
2900
3000
3100
3200
3300
3400
3500
3600
3700
3800
3900
m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
RelativeAbundance
2909.7758
3232.8536
3636.9632
3252.8370
3674.8277
2931.3498
3581.9352
3284.5914
3850.5058
2739.4557
3514.7714
3722.6411
3074.6448
2623.7380
3355.0985
2883.1644
carbonic_140k_inf_003
#
14-31
RT:
0.42-0.93
AV:
18
NL:
9.56E4
T:
FTMS+pNSIsid=1.00 Fullms21000.00@hcd100.00 [500.00-20000.00]
2908.8
2909.0
2909.2
2909.4
2909.6
2909.8
2910.0
2910.2
2910.4
2910.6
2910.8
2911.0
m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
RelativeAbundance
2909.7735
2909.6750
2909.8761
2909.5724
2910.0743
2909.4731
2909.3744
2910.1769
2910.2755
2909.2716
2909.1734
2910.3762
2909.0715
2910.5759
2910.6776
2908.8726
2910.8742
2910.9758
Here the +9 charge state deconvolutes to a mass of
29069.6107 Da. The active protein zinc containing
molecule has a theoretical mass of 29069.6034, relating to
a mass accuracy of 0.25 ppm.
FIGURE 4. Experi
Pyruvate Kinase s
isoforms with bou
bisphosphate (GB
GroEL
#
124-130
RT:
11.29-12.03
AV:
7
NL:
9.95E5
T:
FTMS + p NSI sid=30.00 Full ms2 1000.00@hcd100.
2000
4000
6000
0
10
20
30
40
50
60
70
80
90
100
Relative Abundance
3178.4659 4401.0510
79
6357.0738
FIGURE 5. Spectru
assembly of the 1
PK-qb
#
1
RT:
3.76
AV:
1
NL:
2.00E6
T:
FTMS + p NSI sid=100.00 Full ms [350.00-15000.00]
2000
2500
3000
3500
0
20
40
60
80
100
Relative Abundance
2534.2444
2649.3563
2331.6150
3237.8960
39
GBP
GBP
GBP
monomer
de
PK-qb
#
1
RT:
3.76
AV:
1
NL:
3.99E5
T:
FTMS + p NSI sid=100.00 Full ms [350.00-15000.00]
2000
2200
2400
2600
m/z
0
20
40
60
80
100
Relative Abundance
2534.2444
2428.6827
2649.35
2331.6150
2242.0009
2081.5859
GBP
