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(A)
(D)
Anti-pY antibody
Ion exchange
O
-
O
-
O
-
O=S-O-CH
2
-CH
2
-HN-C-CH
2
-CH
2
-
O
-
O
-
O=S-O-CH
2
-CH
2
-HN-C-CH
2
-CH
2
-
O
O
O=P-O
-
O=P-O
-
O=P-O
-
O
H
3
N
+
-
O
O
G
H
3
N
+
-
pH = 9
pH = 2.7
G
H
2
N-
G
S
-COO
-
T
S
Y
R
S
T
Y
R
-COOH
-COO
-
T
C
Y
R
C
C
NH
C=N
+
H
NH
2
NH
C=NH
NH
2
(B)
(C)
Titanium oxide
IMAC
TiO
2
TiO
2
OH
O
O
N
O
O
O
Fe
3+
O
O=P-O
P
O
O
O
H
3
N
+
-
H
3
N
+
-
G
G
O
S
S
T
-COO
-
Y
R
-COO
-
Y
T
R
C
C
NH
CH
2
=N
+
H
NH
2
NH
CH
2
=N
+
H
NH
2
FIGURE 6
Modes of selecting phosphopeptides. The selectivity of anti-pY antibodies (panel A) is of two types. One selects pY without regard to the
surrounding peptide sequence while the second targets both pY and some of the adjacent amino acid sequence. The IMAC mode of selection (panel B) can
be achieved with Fe
þ
3
as shown, in addition to either Al
þ
3
or Ga
þ
3
on pS, pT, or pY. The selectivity of titanium oxide (panel C) is similar but cross-selects
acidic peptides less. Ion exchanges (panel D) are the least speci
c.
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