Table 1

Phosphorylation site analysis of Bad. Phosphorylated peptides were eluted from the HPLC (Fig 2) were identified by ms/ms mass spectrometry. The amino acid numbers of the peptide in murine Bad are indicated and the phosphorylation site underlined. Results for the release of 32P during Edman degradation cycle sequencing and phospho amino acid analysis of these fractions are also indicated. The percentage of 32P obtained in each HPLC peak, relative to the total 32P eluted in all the peaks is also listed (last column)

Kinase

Peak

Residue Numbers

Peptide Sequence

Phospho-amino acid analysis

Solid Phase cycle no.

Phosphorylation site

% 32P


Pim-1

1

154 – 163

M

    S
DEFEGSFK

Phospho Ser

2

Ser 155

11

2

136 – 146

    S
APPNLWAAQR

Phospho Ser

1

Ser 136

15

3

110 – 131

HS

    S
YPAGTEEDEGMEEELSPFR

Phospho Ser

3

Ser 112

60

4

110 – 131

HS

    S
YPAGTEEDEGMEEELSPFR

Phospho Ser

3

Ser 112

14


Pim-2

3

110 – 131

HS

    S
YPAGTEEDEGMEEELSPFR

Phospho Ser

3

Ser 112

82

4

110 – 131

HS

    S
YPAGTEEDEGMEEELSPFR

Phospho Ser

3

Ser 112

18


Pim-3

1

154 – 163

M

    S
DEFEGSFK

Phospho Ser

2

Ser 155

14

2

136 – 146

    S
APPNLWAAQR

Phospho Ser

1

Ser 136

28

3

110 – 131

HS

    S
YPAGTEEDEGMEEELSPFR

Phospho Ser

3

Ser 112

27

4

110 – 131

HS

    S
YPAGTEEDEGMEEELSPFR

Phospho Ser

3

Ser 112

6

5

unidentified

unidentified

unidentified

N/A

unidentified

13

6

170 – 178

    S
AGTATQMR

Phospho Ser

1

Ser 170

4

7

92 – 107

AA

    T
N
    S
HHGGAGAMETR

Phospho Thr/Ser

3, 5

Thr 94 Ser 96

2

8

170 – 178

    S
AG
    T
ATQMR

Phospho Ser/Thr

1, 4

Ser 170 Thr 173

2

9

unidentified

unidentified

unidentified

N/A

unidentified

3


Macdonald et al. BMC Cell Biology 2006 7:1   doi:10.1186/1471-2121-7-1

Open Data