changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
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After label<strong>in</strong>g, black and red colored spots were cut out from Control group<br />
while the others (green and blue ones) from Bortezomib <strong>applied</strong> group for enzymatic<br />
digestion by us<strong>in</strong>g tryps<strong>in</strong>. Then samples were analyzed by MALDI-TOF-TOF Mass<br />
Spectrometry and mass spectrometric data was compared to the prote<strong>in</strong> database for<br />
sequence matches. If the am<strong>in</strong>o acid sequence of a peptide could be identified, it would<br />
be used to f<strong>in</strong>d out the prote<strong>in</strong> from which it was derived. In addition to this, <strong>in</strong> order to<br />
identify an unknown prote<strong>in</strong> spot, it was necessary for a mass spectrometric analysis to<br />
match more than two peptide’s sequences. In our experiments, prote<strong>in</strong> spot<br />
determ<strong>in</strong>ation was carried out thanks to parameters such as isoelectric po<strong>in</strong>t, molecular<br />
mass, sequence coverage, and mascot mowse score, but the desired peptide matches<br />
were not observed. Although we cannot say that the results obta<strong>in</strong>ed mass spectrometric<br />
analysis are def<strong>in</strong>ite, they are the most probable results because of their parameters and<br />
functions. These results were shown <strong>in</strong> Table 4.2.<br />
Table 4.2 summarizes the 37 differentially expressed prote<strong>in</strong>s that were<br />
characterized. The prote<strong>in</strong>s were identified as follow:<br />
Sixteen prote<strong>in</strong>s were not identified due to some personal and <strong>in</strong>strumental<br />
errors dur<strong>in</strong>g the experiment. These were Spot 1, 13, 16, 17, 19, 20, 21, 24, 26, 28, 30,<br />
31, 32, 33, 36 and 37, namely M1S, S4Cb, K3C, K4C, M6S, M7S, M8S, K6C, S8C,<br />
S10C, M10Sc, M10Sb, M10Sa, K8Cb and K8Ca, respectively.<br />
Spot 2, Caspase-3 is <strong>in</strong>volved <strong>in</strong> the activation cascade of caspases responsible<br />
for apoptosis execution. At the onset of apoptosis, it proteolytically cleaves poly (ADP-<br />
ribose) polymerase (PARP) at a '216-Asp-|-Gly-217' bond and activates sterol<br />
regulatory element b<strong>in</strong>d<strong>in</strong>g prote<strong>in</strong>s (SREBPs) between the basic helix-loop-helix<br />
leuc<strong>in</strong>e zipper doma<strong>in</strong> and the membrane attachment doma<strong>in</strong>. Additionally, it cleaves<br />
and activates caspase-6, -7 and -9. Caspase-3 found <strong>in</strong> many cell l<strong>in</strong>es, highest<br />
expression <strong>in</strong> cells of the immune system. Because of its function and role <strong>in</strong> apoptosis,<br />
after apply<strong>in</strong>g Bortezomib (17 nM) to MM U-266 cells, the <strong>in</strong>creas<strong>in</strong>g expression level<br />
of this prote<strong>in</strong> was expected results.<br />
Spot 3 were signed to uncharacterized prote<strong>in</strong> C7orf46. Although its function<br />
does not know exactly, it is thought that this prote<strong>in</strong> may be <strong>in</strong>volved <strong>in</strong> alternative<br />
splic<strong>in</strong>g.<br />
Spot 18, C-type lect<strong>in</strong> doma<strong>in</strong> family 5 member A has a function as a positive<br />
regulator of osteoclastogenesis. Thus, it was upregulated after the cells exposed to<br />
Bortezomib.<br />
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