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2014 ; 60
(4
): 1314-23
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Shotgun proteomics: identification of unique protein profiles of apoptotic bodies
from biliary epithelial cells
#MMPMID24841946
Lleo A
; Zhang W
; McDonald WH
; Seeley EH
; Leung PS
; Coppel RL
; Ansari AA
; Adams DH
; Afford S
; Invernizzi P
; Gershwin ME
Hepatology
2014[Oct]; 60
(4
): 1314-23
PMID24841946
show ga
Shotgun proteomics is a powerful analytic method to characterize complex protein
mixtures in combination with multidimensional liquid chromatography-tandem mass
spectrometry (LC-MS/MS). We used this platform for proteomic characterization of
apoptotic bodies in an effort to define the complex protein mixtures found in
primary cultures of human intrahepatic biliary epithelial cells (HiBEC), human
renal proximal tubular epithelial cells, human bronchial epithelial cells,
isolated intrahepatic biliary epithelial cells from explanted primary biliary
cirrhosis (PBC), and control liver using a total of 24 individual samples.
Further, as additional controls and for purposes of comparison, proteomic
signatures were also obtained from intact cells and apoptotic bodies. The data
obtained from LC-MS/MS, combined with database searches and protein assembly
algorithms, allowed us to address significant differences in protein spectral
counts and identify unique pathways that may be a component of the induction of
the signature inflammatory cytokine response against BECs, including the Notch
signaling pathway, interleukin (IL)8, IL6, CXCR2, and integrin signaling. Indeed,
there are 11 proteins that localize specifically to apoptotic bodies of HiBEC and
eight proteins that were specifically absent in HiBEC apoptotic bodies.
CONCLUSION: Proteomic analysis of BECs from PBC liver compared to normal liver
are significantly different, suggesting that an immunological attack affects the
repertoire of proteins expressed and that such cells should be thought of as
living in an environment undergoing continuous selection secondary to an innate
and adaptive immune response, reflecting an almost "Darwinian" bias.
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