In:
PROTEOMICS, Wiley, Vol. 6, No. 2 ( 2006-01), p. 614-622
Abstract:
Proteome analysis of Jurkat T cells induced to undergo apoptosis by CD95 (Fas/Apo‐1) treatment was performed to identify modified proteins. We used stable isotope labeling with amino acids in cell culture (SILAC) using leucine to identify proteins of apoptotic and control Jurkat T cells by 2‐DE and MALDI‐MS. Out of 224 spots analyzed, we quantified 213 spots with 3.5 leucine‐containing peptide pairs on average; 28 proteins with a relative abundance of higher than 1.5 were found. Five new modified proteins including calcyclin binding protein, cytosolic acyl coenzyme A thioester hydrolase, heterogeneous ribonucleoprotein M, replication factor C 37‐kDa subunit, and tropomyosin 4 chain were identified as being modified in response to apoptosis. In comparison to differential proteome analysis via silver‐stained 2‐D gels and PMF of total Jurkat T cell lysates, 15 additional apoptosis‐modified proteins were identified though 8 proteins were not found. The described approach using SILAC instead of silver staining for relative quantification was simpler to perform regarding the number of required 2‐D gels, that cumbersome gel comparisons were avoided, and more apoptosis‐modified proteins were identified, but with a higher demand on data interpretation of the mass spectra obtained.
Type of Medium:
Online Resource
ISSN:
1615-9853
,
1615-9861
DOI:
10.1002/pmic.200500120
Language:
English
Publisher:
Wiley
Publication Date:
2006
detail.hit.zdb_id:
2037674-1
detail.hit.zdb_id:
2032093-0
SSG:
12
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