58
' ~~~O~~~~~~~~~N~O~~~O
I
• • • • • • • • • • • • • • • • • • • •
G' ~.~"~"~G~.~~~~~~~~DO
n: ~~~m~~;~~::~:~~~~~~~
I
. . . . . . . . . . . .
. . . ... ~ .. " " .. . . . . .. . . ... . . ........
QI ~~~~~"'N"'~~ O "'~ ~ "' ON"~ ~
U I " ~~ ~ ~ O~ ~~~"' N ~~~ ~ ~ "O ~
"'I ~~.~~~.~.~~~ "~ "' " ~. ~ •
., I
• • • • • • • • • • • • • • • • • • • •
QI OOOOOOOOOOOOOOOO O OOO
" I G~~~mN"'~"~~ND"''''~~'''~.
t! :~E~~Es9~~E:~!~s:=S~
U I • • • • • • • • • • • • • • • • • • ••
MI NNNNNNNNNHN N NN"NN " NN
-------------------- NNnn ......... NN~ ...... O~nC ...... m~
oooooooooooo~+·ooooooo
~~!!~~~~~~~~_~tt~~t~
" I NOOHNNNNNNNN.~~N~~~~
.,. I
•
• • • • • • • • • • • • • • •
•
•
• •
~: ~~~~2~9S9999~~~~~~8o
!, O~GOOOOOODOO ... " . . . . . NNN
i. "
................................................ ...
I ... N".~~~N~O .... ~~N~~ ... M~
.,
...... NNNNN""".~.
,
~
't ~
~
~
~
~;I
~..,.
~ ..
:i!l~1
~
~
"' 1::1
01 .. 1
•• ""I
0 _
·0
o •
~~
'N
o •
. . .,
NN
....
N"
"N
J. R. YATES et al.
.5
,.D
S
0.0
0
e Q)
~
- -
r/)
Q)
Q
> ~
Q)
~
- >1
~
t
Q
~
.- ~
00
00
~
~
.- 00
~
-~ a
s:l
0
.- .- .s
~
:=E
Fig. 4.2. SEQUEST output from the analysis of the mutant protein responsible for sickle cell anemia.
Tandem mass spectra were acquired from both the normal and mutant protein after proteolytic
digestion of separate aliquots using the proteases trypsin, endoproteinase Glu-C, subtilisin, and chymotrypsin. Approximately 99 % of the sequence was represented by tandem mass spectra after LCI
MS/MS analysis. Tandem mass spectra acquired from the normal protein were subtracted from that
of the mutant protein. The remaining spectra were then subjected to a database search using the
SEQUEST-SNP program