Table 4.1. Magnified Segments of Differentiated Prote<strong>in</strong> Spots (cont.) BORTEZOMIB CONTROL Spot Name Spot No S5C 22 K5C 23 K6C 24 S6C 25 K7C 26 Y5S S8C S9C S10C M10Sc M10Sb M10Sa M9S Y4S K8Cb K8Ca 27 28 29 30 31 32 33 34 35 36 37 75
After label<strong>in</strong>g, black and red colored spots were cut out from Control group while the others (green and blue ones) from Bortezomib <strong>applied</strong> group for enzymatic digestion by us<strong>in</strong>g tryps<strong>in</strong>. Then samples were analyzed by MALDI-TOF-TOF Mass Spectrometry and mass spectrometric data was compared to the prote<strong>in</strong> database for sequence matches. If the am<strong>in</strong>o acid sequence of a peptide could be identified, it would 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 identify an unknown prote<strong>in</strong> spot, it was necessary for a mass spectrometric analysis to match more than two peptide’s sequences. In our experiments, prote<strong>in</strong> spot determ<strong>in</strong>ation was carried out thanks to parameters such as isoelectric po<strong>in</strong>t, molecular mass, sequence coverage, and mascot mowse score, but the desired peptide matches were not observed. Although we cannot say that the results obta<strong>in</strong>ed mass spectrometric analysis are def<strong>in</strong>ite, they are the most probable results because of their parameters and functions. These results were shown <strong>in</strong> Table 4.2. Table 4.2 summarizes the 37 differentially expressed prote<strong>in</strong>s that were characterized. The prote<strong>in</strong>s were identified as follow: Sixteen prote<strong>in</strong>s were not identified due to some personal and <strong>in</strong>strumental errors dur<strong>in</strong>g the experiment. These were Spot 1, 13, 16, 17, 19, 20, 21, 24, 26, 28, 30, 31, 32, 33, 36 and 37, namely M1S, S4Cb, K3C, K4C, M6S, M7S, M8S, K6C, S8C, S10C, M10Sc, M10Sb, M10Sa, K8Cb and K8Ca, respectively. Spot 2, Caspase-3 is <strong>in</strong>volved <strong>in</strong> the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis, it proteolytically cleaves poly (ADP- ribose) polymerase (PARP) at a '216-Asp-|-Gly-217' bond and activates sterol regulatory element b<strong>in</strong>d<strong>in</strong>g prote<strong>in</strong>s (SREBPs) between the basic helix-loop-helix leuc<strong>in</strong>e zipper doma<strong>in</strong> and the membrane attachment doma<strong>in</strong>. Additionally, it cleaves and activates caspase-6, -7 and -9. Caspase-3 found <strong>in</strong> many cell l<strong>in</strong>es, highest expression <strong>in</strong> cells of the immune system. Because of its function and role <strong>in</strong> apoptosis, 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 of this prote<strong>in</strong> was expected results. Spot 3 were signed to uncharacterized prote<strong>in</strong> C7orf46. Although its function does not know exactly, it is thought that this prote<strong>in</strong> may be <strong>in</strong>volved <strong>in</strong> alternative splic<strong>in</strong>g. Spot 18, C-type lect<strong>in</strong> doma<strong>in</strong> family 5 member A has a function as a positive regulator of osteoclastogenesis. Thus, it was upregulated after the cells exposed to Bortezomib. 76
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CHANGES IN PROTEIN PROFILES IN BORT
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ACKNOWLEDGEMENTS First of all, with
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ÖZET BORTEZOMĠB UYGULANAN MULTĠP
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2.1.4.1.2. Two-Dimensional Gel Elec
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LIST OF FIGURES Figure Page Figure
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LIST OF TABLES Table Page Table 1.1
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When normal cells in the body begin
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Figure 1.2. Development of Cancer S
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the myeloma cells does not help fig
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A rare form of MM called Nonsecreto
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1.2.3. Risk Factors for MM A risk f
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of the mineralizations by adjusting
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To measure the levels of red cells,
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The levels of blood urea nitrogen (
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By using the system introduced by D
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Figure 1.10. Ubiquitin-Proteasome P
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