Background
Results
Subcellular fractionation
Electron micrographs of isolated vesicles
2D-DIGE of enriched secretory granules
Peptide mass fingerprinting of selected spots
spot # | protein name | Vol. ratio | NCBI Acc.No. | MW | pI | identified peptides | MASCOT score |
---|---|---|---|---|---|---|---|
1 | MHC class II alpha chain | -2.13 | gi|3212400 | 20548 | 4.9 | 3 | 113 |
2 | capping protein (actin filament), gelsolin-like | -3.03 | gi|63252913 | 38760 | 5.8 | 9 | 334 |
3 | chaperonin containing TCP1, subunit 8 | 2.63 | gi|48762932 | 60153 | 5.4 | 5 | 183 |
4 | capping protein (actin filament), gelsolin-like | 8.98 | gi|55597035 | 38779 | 5.9 | 9 | 368 |
Similarity of individual SL preparations
Discussion
Conclusion
Methods
Cells
Subcellular fractionation
fraction | Exp. 1 | Exp. 2 | Exp. 3 | Exp. 4 | mean | SD | Volume | protein |
---|---|---|---|---|---|---|---|---|
[mg/ml] | [mg/ml] | [mg/ml] | [mg/ml] | [mg/ml] | [μl] | [μg] | ||
homogenate
| 3.891 | 3.222 | 3.532 | 3.684 |
3.582
| 0.282 |
2700
|
9671.4
|
enriched organelles (input)
| 0.745 | 0.651 | 0.729 | 0.865 |
0.747
| 0.088 |
1800
|
1345.3
|
fraction 1
| 0.349 | 0.303 | 0.347 | 0.296 |
0.324
| 0.028 |
200
|
64.7
|
fraction 2
| 0.430 | 0.468 | 0.306 | 0.389 |
0.398
| 0.069 |
400
|
159.3
|
fraction 3
| 0.764 | 0.909 | 1.004 | 0.879 |
0.889
| 0.099 |
300
|
266.7
|
fraction 4
| 0.526 | 0.548 | 0.480 | 0.405 |
0.490
| 0.063 |
300
|
147.0
|
fraction 5
| 0.925 | 0.882 | 0.959 | 0.852 |
0.905
| 0.047 |
200
|
180.9
|
fraction 6
| 0.384 | 0.431 | 0.352 | 0.349 |
0.379
| 0.038 |
200
|
75.8
|
recovery
|
894.4
|