Protein Signatures of Leukemic Cells
EMSL Project ID
2622
Abstract
The objective of this project is to determine whether differences in protein expression in acute promyelocytic leukemia with a t(15;17) segregate with treatment outcome. The strategy to identify protein markers is to identify proteins in leukemias with known treatment outcome by identifying proteins of low and high abundance proteins separated on SDS PAGE gels. Initial studies to optimize technical strategies are carried out with the APL cell line NB4, which can be induced to differentiate to granulocytes by treatment with all-trans retinoic acid. We will determine whether this strategy allows detection of low abundance proteins that are differentially expressed in differentiated and undifferentiated cells, and enhance proteome information from clinical specimens. One band from a 1-D gel of cellular lysates (differentiated and undifferentiated, in duplicate) will be trypsin digested and subjected to HPLC using a 50 micron ID column followed by LCQ ion trap MS/MS analysis for protein identifications. We anticipate that the information allow us to identify both low and high abundance proteins, effectively increasing the dynamic range of proteome representation on the gel. These data will establish the nature and dynamic range of comparative proteomics using 1D gel separation followed by LC-MS/MS, particularly for low abundance proteins. Demonstration of reproducible detection of low abundance proteins using this method would indicate feasibility of using this strategy to compare low abundance profiles among specimens and warrant wider-scale development of this approach.
Project Details
Project type
Exploratory Research
Start Date
2002-09-25
End Date
2003-03-10
Status
Closed
Released Data Link
Team
Principal Investigator