Eritrositlerde L-sistein amino asit exportuna hücre içi ve dışı redoks durumunun etkisi
The effect of intracellular and extracellular redox status on L-cysteine export in erythrocytes
Yazar: CEYLAN USLU
Danışman: DOÇ.DR. DENİZ YILDIZ
Mustafa Kemal Üniversitesi / Fen Bilimleri Enstitüsü / Genel Biyoloji Ana Bilim Dalı
Derece: Yüksek Lisans, 2006
Konu: Biyokimya = Biochemistry ; Biyoloji = Biology

Özet:
1ÖZET(5Ã¸7526Ã¸7/(5'(/6Ã¸67(Ã¸1$0Ã¸12$6Ã¸7(;325781$+h&5(Ã¸dÃ¸9(',Ã¹,5('2.6'85808181(7.Ã¸6Ã¸Glutatyon, antioksidan savunmada önemli bir rol oynar. Bu nedenle glutatyon,eritrositlerde yüksek miktarlarda bulunur. L-sistein amino asidi, eritrositlerde glutatyonVHQWH]L LoLQ JHUHNPHNWHGLU %X oDOÃÃºPDQÃQ DPDFÃ KÂFUH LoL YH GÃÃºÃ redoks durumunun,eritrositlerde L-sistein HNVSRUWXQD HWNLVLQL LQFHOHPHNWLU dDOÃÃºPDGD HULWURVLWOHUL IDUNOÃkonsantrasyonlardaki L-sistein DPLQR DVLGLQH PDUX] EÃUDNÃOPÃÃº YH VRQUD GDHULWURVLWOHUGHNL VHUEHVW Â±6+ NRQVDQWUDV\RQXQX |OoÂOPÂÃºWÂU $\QÃ \|QWHPOH PXDPHOHedilen eritrositler, sonraki L-VLVWHLQ oÃNÃÃºÃ oDOÃÃºPDODUÃQGD NXOODQÃOPÃÃºWÃU $\QÃ ]DPDQGDÃVÃQÃQ EX JLULÃºoÃNÃÃºD RODQ HWNLVL GHÃ·HUOHQGLULOPLÃºWLU Ã¸ON RODUDN ELU glutatyon tüketicisiolan 1-chloro-2,4-dinitrobenzene ile muamele edilen eritrositlerde, buthionineVXOIR[LPLQHYDUOÃÃ·ÃQGDYH\RNOXÃ·XQGD/VLVWHLQoÃNÃÃºÃQÃQQDVÃOHWNLOHQGLÃ·LEHOLUOHQPLÃºWLUL-VLVWHLQ JLULÃºL oDOÃÃºPDODUÃPÃ]GD SOD]PDGD /VLVWHLQ NRQVDQWUDV\RQX DUWWÃÃ·ÃQGDHULWURVLWOHULQ EX DUWÃÃºD FHYDS YHUHUHN /sistein DPLQR DVLGLQL KÂFUH LoLQH DOGÃÃ·ÃJ|VWHULOPLÃºWLU (ULWURVLWOHUGHNL VHUEHVW Â±6+ NRQVDQWUDV\RQX P0 /sisteinX\JXODQGÃÃ·ÃQGDVDDWWHÂP0Â¶D \ÂNVHOLUNHQEXNRQVDQWUDV\RQ P0 /VLVWHLQ X\JXODQGÃÃ·ÃQGD Â P0Â¶ D \ÂNVHOPLÃºWLU (ULWURVLWOHUGH VHUEHVW Â±6+NRQVDQWUDV\RQODUÃ P0 /VLVWHLQ X\JXODQGÃÃ·ÃQGD GDNLNDGD Â P0Â¶ D\ÂNVHOLUNHQVDDWWHEXNRQVDQWUDV\RQÂP0Â¶D\ÂNVHOPLÃºWLU$\QÃ]DPDQGDL-sistein DPLQR DVLGLQLQ oÃNÃÃºÃQÃQ GD ]DPDQ YH NRQVDQWUDV\RQD EDÃ·OÃ ROGXÃ·XEHOLUOHQPLÃºWLU <ÂNVHN PLNWDUGD /VLVWHLQOH PXDPHOH HGLOPLÃº HULWURVLWOHUGH \ÂNVHNmiktarda L-VLVWHLQoÃNÃÃºÃJ|UÂOÂUP0/sisteinle muamele edilen eritrositlerde, hücreGÃÃºÃVHUEHVWÂ±6+NRQVDQWUDV\RQXVDDWWHÂP0Â¶D\ÂNVHOLUNHQP0/sisteinle muamele edilen eritrositlerde bu konsantrasyon 1,014 Â± 0,002 mM' a\ÂNVHOPLÃºWLU Â& VÃFDNOÃN HULWURVLWOHUH /VLVWHLQ JLULÃºLQL WHVW HGLOHQ EÂWÂQkonsantrasyonlarda, önemli derecede LQKLEH HWPLÃºWLU 6ÃFDNOÃN DUWÃÃºÃ EX JLULÃº ROD\ÃQÃ\HQLGHQ GÂ]HOWPHNWHGLU %HQ]HU VRQXoODU oÃNÃÃº oDOÃÃºPDODUÃ LOH GH HOGH HGLOPLÃºWLU6RQXoODUÃPÃ] D\QÃ ]DPDQGD /VLVWHLQLQ LoHUL YH GÃÃºDUÃ transportunun, eritrositinRNVLGDWLI GXUXPXQGDQ HWNLOHQGLÃ·LQL J|VWHUPLÃºWLU *OXWDW\RQ WÂNHWLOGLÃ·LQGH YH glutatyonVHQWH]L HQJHOOHQGLÃ·LQGH /VLVWHLQLQ KÂFUH\H JLULÃºoÃNÃÃºÃ |QHPOL GHUHFHGH D]DOPÃÃºWÃUEritrositler, plazma UHGRNV GXUXPXQXQ GHYDPÃ YH KÂFUH LoL JOXWDW\RQXQ EHOLUOHGLÃ·L /VLVWHLQoÃNÃÃºRUDQÃQGDUROR\QDU2006, 46 sayfaAnahtar Kelimeler: Eritrosit, 6LVWHLQoÃNÃÃºÃRedoks dengesi.

Abstract:
ABSTRACTTHE EFFECT OF INTRACELLULAR AND EXTRACELLULAR REDOX67$78621/&<67(Ã¸1((;3257,1(5<7+52&<7(6Glutathione has an important role in antioxidant defense. For this reason,glutathione is found at rather high amounts in erythrocytes. L-cysteine is required forglutathione synthesis in erythrocytes. The objective of this study was to invastigate theeffect of intracellular and extracellular redoks status on L-cysteine exports inerythrocytes. In the present study, we exposed the erythrocytes to differentconcentrations of L-cysteine and then measured the intracellular free -SHconcentrations. The erythrocytes treated in the same manner were later utilized for thecysteine efflux studies. The effect of temperature on the influx and the efflux processeswere also evaluated. We also determined the rate of L-cysteine efflux in the presenceand absence of Buthionine Sulfoximine in erythrocytes that are pretreated with 1-chloro-2, 4-dinitrobenzene, a glutathione depleter.Our L-cysteine influx studies demonstrated that erythrocytes can respond toincreases in L-cysteine concentration in the extracellular media and influx L-cysteine ina concentration-dependent manner. Free -SH concentration in erythrocytes treated with1 mM L-cysteine reached to 1,64 Â± 0,06 mM in 1 hour whereas this concentrationreached to 4,30 Â± 0,01 mM in 10 mM L-cysteine treated erythrocytes. Free -SHconcentration in erythrocytes treated with 10 mM L-cysteine reach to 2,48 Â± 0,09 mMin 10 minute whereas this concentration, when treated 10 mM L-cysteine in 1 hour,reach to 4,30 Â± 0,01 mM. The L-cysteine efflux is also determined to be time andconcentration-dependent. Erythrocytes that are pretreated with higher L-cysteineconcentrations displayed a higher efflux process. Outside concentration of free -SH in 1mM L-cysteine pretreated erythrocytes, reach to 0,200 Â± 0,005 mM in 1hour, whereasthis concentration reach to 1,014 Â± 0,002 mM with 10 mM L-cysteine pretreatederythrocytes. The influx process is significantly inhibited +4 ºC at all of theconcentration tested. Increasing of the temperature later recovered the influx process.Similar results were obtained with the efflux studies. Our results also indicate that rateof inward and outward transport of L-cysteine is affected by the oxidative status oferythrocytes. When glutathione is depleted and glutathione synthesis is blocked, L-cysteine uptake and the efflux processes are significantly decreased. Erythrocytes play arole in regulation the plasma redoks status and intracellular level of glutathionedetermines the rate of the L-cysteine efflux.2006, 46 PagesKeywords: Erythrocytes, Cysteine efflux, Redox homeostasis