Table 3.

Quantitating the effect of Sch.414319 on phosphoproteins detected in human mononuclear cell preparations

Protein reference no.	Protein pI	Protein MWt	Sch.414319 Difference	HU210 Difference
23	8.34	64 541	2616	—
35	6.62	56 000	301	—
44	6.6	47 000	554	—
53	6.81	40 833	6559	—
71	4.9	25 814	4879	1423
78	5.24	23 173	2138	—
1	ND	ND	−91	−97
2	5.28	194 711	−88	—
5	5.67	156 000	−89	—
16	5.77	71 000	−98	—	L-plastin
52	5.84	41 000	−78	—
57	5.87	37 616	−97	—
61	5.15	35 000	−69	—	14-3-3-ɛ
69	5.46	28 647	—	−91
70	4.74	26 000	−99	—
72	6.24	25 057	−72	—
76	5.94	23 000	−70	—
77	5.13	23 424	−96	—
86	4.83	20 000	−78	—
96	ND	ND	—	−91
7	5.61	119 000	−52	137

Abbreviations: MWt, molecualr weight; ND, not done.

Cell-free extracts from [³²P] orthophosphoric acid-labelled human peripheral blood mononuclear cells plus/minus drug treatment (10 nM, 37 °C, 60 min) were analysed by two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis, followed by autoradiography. Resulting films were digitally analysed and quantitated by Kendrick Laboratories (Madison, WI, USA). Differences in film darkening were quantitated according to the following formula:

Difference=(1−spot% sample × /spot % sample ref.)(−100).

Proteins in grey were selected for peptide sequencing by MALDI-MS (Protein Chemistry Core Facility, Columbia University).