{"id":95,"date":"2020-07-22T21:52:00","date_gmt":"2020-07-22T18:52:00","guid":{"rendered":"https:\/\/www.ortaakarsu.net\/?p=95"},"modified":"2021-05-07T21:10:57","modified_gmt":"2021-05-07T18:10:57","slug":"glutatyon-reduktaz-enzim-aktivitesi","status":"publish","type":"post","link":"https:\/\/www.ortaakarsu.net\/?p=95","title":{"rendered":"Glutatyon Red\u00fcktaz Enzim Aktivitesi"},"content":{"rendered":"\n

Glutatyon red\u00fcktaz aktivitesi ile ilgili bu yaz\u0131daki ligandlar\u0131n sentezi, yap\u0131 ayd\u0131nlatmas\u0131, in vitro \u00e7al\u0131\u015fmas\u0131 Prof. Dr. Nurcan Karacan<\/a> ve Dr. \u00d6\u011fr. \u00dcyesi Turgay TUN\u00c7<\/a>‘a aittir<\/strong>. Bu ligandlarla yap\u0131lan docking \u00e7al\u0131\u015fmas\u0131 ve sonu\u00e7lar\u0131n yorumlanmas\u0131 Prof. Dr. Nurcan Karacan<\/a> ve Ahmet B. Ortaakarsu taraf\u0131ndan yap\u0131lm\u0131\u015ft\u0131r.<\/p>\n\n\n\n

1. AMA\u00c7<\/strong><\/strong><\/h2>\n\n\n\n

Bu \u00e7al\u0131\u015fmada, daha \u00f6nceden sentezlenmi\u015f ve in vitro<\/em> glutatyon red\u00fcktaz inhibitor aktiviteleri denel olarak belirlenmi\u015f yeni Schiff bazlar\u0131n\u0131n (1-4)<\/strong> molek\u00fcler yerle\u015ftirme (docking) \u00e7al\u0131\u015fmalar\u0131 yap\u0131lacakt\u0131r.<\/p>\n\n\n\n

In vitro<\/em> \u00e7al\u0131\u015fmalar Schiff bazlar\u0131n\u0131n (4 <\/strong>hari\u00e7)<\/strong> glutatyon red\u00fcktaz (GR) enzimine kar\u015f\u0131 inhibitor olarak davrand\u0131\u011f\u0131n\u0131 g\u00f6stermi\u015ftir. Schiff bazlar\u0131n\u0131n inhibisyon etkileri simulasyon hesaplamalar\u0131 ile analiz edilecektir. B\u00f6ylece, ligandlar\u0131n GR enzimine ba\u011flanma yerleri ve ba\u011flanma afiniteleri (G skor) teorik olarak belirlenecektir.<\/p>\n\n\n\n

Hesaplamalar farkl\u0131 canl\u0131lardaki GR enzimleri (homo sapiens, Saccharomyces cerevisiae, Plasmodium falciparum<\/em>) \u00fczerinde tekrarlanacak, b\u00f6ylece yeni Schiff bazlar\u0131n\u0131n canl\u0131lar \u00fczerindeki olas\u0131 etkileri \u00f6ng\u00f6r\u00fclebilir hale gelecektir. Sonu\u00e7lar sonraki in vitro<\/em> ve in vivo<\/em> \u00e7al\u0131\u015fmalar\u0131 i\u00e7in \u00f6nemli bir zemin haz\u0131rlayacakt\u0131r.<\/p>\n\n\n\n

Denel veriler 4<\/strong> nolu bir Schiff baz\u0131n\u0131n enzim aktivat\u00f6r\u00fc olarak davrand\u0131\u011f\u0131n\u0131 g\u00f6stermi\u015ftir. Bu farkl\u0131l\u0131\u011f\u0131n nedeni docking \u00e7al\u0131\u015fmalar\u0131 ile ayd\u0131nlat\u0131lacakt\u0131r. Hesaplamalar ile GR enziminde allosterik merkezin yeri tam olarak belirlenmeye \u00e7al\u0131\u015f\u0131lacakt\u0131r.<\/p>\n\n\n\n

2. <\/strong>GLUTATYON RED\u00dcKTAZ VE \u0130\u015eLEV\u0130(KURAMSAL TEMELLER)<\/h2>\n\n\n\n

2.1 Glutatyon Red\u00fcktaz Enzimi<\/h3>\n\n\n\n

Glutatyon red\u00fcktaz (GR, EC 1.8.1.7), prokaryot ve \u00f6karyot h\u00fccrelerde bulunan NADPH’ye ba\u011fl\u0131 oksidored\u00fcktaz bir enzimdir; 2×52 kDa’luk homodimerik flavoproteindir, oksidatif strese kar\u015f\u0131 h\u00fccreyi korur [Dolphin ve ark. (1989)]. Bu antioksidan enzim, glutatyon dis\u00fclfitin (GSSG, y\u00fckseltgenmi\u015f glutatyon) glutatyona (GSH, indirgenmi\u015f glutatyon) indirgenmesini katalizler. GSSG, oksidatif stress ortam\u0131nda, GSH’nin y\u00fckseltgenmesi ile ortaya \u00e7\u0131kar, bu esnada di\u011fer reaktif oksijen t\u00fcrleri (ROS) de olu\u015fur. Sitoplazmada GSH\/GSSG oranlar\u0131 30:1 ila 300:1 aras\u0131nda de\u011fi\u015fir [Jones (2002); Zhao ve ark.,(2009)]. Y\u00fcksek GSH deri\u015fimi nedeniyle, GSH\/GSSG, h\u00fccrenin ana redoks tamponu olarak kabul edilir, GSH\/GSSG oran\u0131 oksidatif stres ko\u015fullar\u0131nda azal\u0131r.<\/p>\n\n\n\n

\"\"<\/figure><\/div>\n\n\n\n

GR inhibit\u00f6rleri, antimalarya, antikanser ilac\u0131 ve kemoterapi\/ radyoterapide duyarl\u0131l\u0131\u011f\u0131 artt\u0131rma ajan\u0131 olarak kullan\u0131lmaktad\u0131rlar. S\u0131tma tedavisi i\u00e7in GR enzimini hedef alan ila\u00e7lar ile ilgili pek \u00e7ok \u00e7al\u0131\u015fma mevcuttur [Bauer ve ark (2006); Becker ve ark.(2003); Biot ve ark.(2003); Davioud ve ark. (2001)]. Antikanser ilac\u0131 olarak kullan\u0131lan GR inhibit\u00f6rleri oksidatif stresi art\u0131rarak t\u00fcm\u00f6rlerin \u00e7o\u011falmas\u0131n\u0131 engeller. GSSG art\u0131\u015f\u0131 h\u00fccre fonksiyonlar\u0131n\u0131 bozar [Karplus ve ark. (1988)]. Radyoterapinin kanser h\u00fccrelerini \u00f6ld\u00fcrmesindeki etken ROS \u00fcretimidir. GR inhibit\u00f6rleri oksidatif stresi artt\u0131r\u0131larak radyoterapiye destek sa\u011flamaktad\u0131r.<\/p>\n\n\n\n

Enzimler ile ilgili daha \u00e7ok bilgi almak i\u00e7in Enzimler ve Genel \u00d6zellikleri<\/a> yaz\u0131s\u0131n\u0131 okuyunuz.<\/p>\n\n\n\n

Proje kapsam\u0131nda kullan\u0131lan GR enzimlerinin 3D-kristal geometrileri ile aktif b\u00f6lge etkile\u015fim yap\u0131lar\u0131 a\u015fa\u011f\u0131da verilmektedir.<\/p>\n\n\n\n

\"\"
\u015eekil 2.1a<\/strong> (PDB: 3DK8<\/a>) insan eritrosit<\/em> enzimi ile ligand\u0131n\u0131n aktif b\u00f6lge ile etkile\u015fim yap\u0131s\u0131 ve kofakt\u00f6r\u00fc
DOI: <\/strong>10.2210\/pdb3DK8\/pdb<\/a><\/figcaption><\/figure><\/div>\n\n\n\n
\"\"
\u015eekil 2.1b<\/strong> (PDB: 1BWC<\/a>) insan eritrosit<\/em> glutatyon red\u00fcktaz enzimi ile ligand\u0131n\u0131n aktif b\u00f6lge ile etkile\u015fim yap\u0131s\u0131 ve kofakt\u00f6r\u00fc
DOI: <\/strong>10.2210\/pdb1BWC\/pdb<\/a>
<\/figcaption><\/figure><\/div>\n\n\n\n
\"https:\/\/www.ortaakarsu.net\/?p=6392\"
\u015eekil 2.1c<\/strong> (PDB: 2GH5<\/a>) insan eritrosit<\/em> enzimi ile ligand\u0131n\u0131n aktif b\u00f6lge ile etkile\u015fim yap\u0131s\u0131 ve kofakt\u00f6r\u00fc
DOI: <\/strong>10.2210\/pdb2GH5\/pdb<\/a><\/figcaption><\/figure><\/div>\n\n\n\n
\"bir
\u015eekil 2.1d <\/strong>(PDB: 2HQM<\/a>) Saccharomyces cerevisiae<\/em>‘da Glutatyon Red\u00fcktaz enzimi ve kofakt\u00f6r\u00fc
DOI: <\/strong>10.2210\/pdb2HQM\/pdb<\/a><\/figcaption><\/figure><\/div>\n\n\n\n
\"bir
\u015eekil 2.1e<\/strong> (PDB: 1ONF<\/a>) Plasmodium Falciparum<\/em>‘da Glutatyon Red\u00fcktaz enzimi ve kofakt\u00f6r\u00fc
DOI: <\/strong>10.2210\/pdb1ONF\/pdb<\/a><\/figcaption><\/figure><\/div>\n\n\n\n

2.2 Schiff Bazlar\u0131<\/strong> ve Glutatyon Red\u00fcktaz<\/h3>\n\n\n\n

\u00d6nce 2,6-dis\u00fclfanilpiridin-2,2\u2019-dianilin ba\u015flang\u0131\u00e7 bile\u015fi\u011fi (P) sentezlenmi\u015f ve yap\u0131s\u0131 X-\u0131\u015f\u0131nlar\u0131 difraksiyon y\u00f6ntemi ile ayd\u0131nlat\u0131lm\u0131\u015ft\u0131r (\u015eekil 2.2a).<\/p>\n\n\n\n

\"glutatyon
\u015eekil 2.2a 2,6-dis\u00fclfanilpiridin-2,2\u2019-dianilin bile\u015fi\u011finin 3D yap\u0131s\u0131<\/figcaption><\/figure><\/div>\n\n\n\n
\n

Bu ba\u015flang\u0131\u00e7 bile\u015fi\u011fine \u00e7e\u015fitli aldehitler kat\u0131larak 4 farkl\u0131 Schiff Baz\u0131 (1-4)<\/strong> sentezlenmi\u015f ve yap\u0131lar\u0131 spektrofotometrik y\u00f6ntemlerle ayd\u0131nlat\u0131lm\u0131\u015ft\u0131r (\u015eekil 2.2b). Bile\u015fiklerin isimleri a\u015fa\u011f\u0131da verilmektedir.<\/p>\n\n\n\n

(1) 2,6-Bis (2-{(Z)-[(2-s\u00fclfanilfenil)iminometil}fenol)piridin<\/p>\n\n\n\n

(2)2,6-Bis(2-{[(1Z,2E)-2-(hidroksimino)-1-metilpropiliden]amino}-benzentiyol) piridin<\/p>\n\n\n\n

(3) 2,6-Bis (2-{[(1Z)-tiyofen-2-il-metiliden]amino}benzentiyol)piridin<\/p>\n\n\n\n

(4) 2,6-Bis (2-{[(1Z)-1H-imidazol-4-il-metiliden]amino}benzentiyol)piridin<\/p>\n<\/div><\/div>\n\n\n\n

\"glutatyon
\u015eekil 2.2b Doking \u00e7al\u0131\u015fmas\u0131 yap\u0131lan bile\u015fiklerin a\u00e7\u0131k yap\u0131s\u0131<\/figcaption><\/figure><\/div>\n\n\n\n
\n

Bu bile\u015fiklerin ekmek mayas\u0131 ve insan eritrositinden izole edilmi\u015f GR enzimleri \u00fczerindeki etkileri in vitro<\/em> \u00e7al\u0131\u015f\u0131lm\u0131\u015f ve IC50 <\/sub>de\u011ferleri \u00c7izelge 2.2 de verilmi\u015ftir.<\/p>\n\n\n\n

\u00c7izelge 2.2. Bile\u015fiklerin glutatyon red\u00fcktaz aktivite IC50<\/sub> de\u011ferleri<\/p>\n\n\n\n

Bile\u015fikler<\/td>Ekmek mayas\u0131 IC50<\/sub> (\u00b5M)<\/td>\u0130nsan eritrositi IC50<\/sub> (\u00b5M)<\/td><\/tr>
P<\/td>135.62\u00b11.00<\/td>143.14\u00b11.07<\/td><\/tr>
(1)<\/td>55.23\u00b10.27<\/td>66.87\u00b10.36<\/td><\/tr>
(2)<\/td>52.42\u00b10.71<\/td>63.56\u00b10.25<\/td><\/tr>
(3)<\/td>50.09\u00b10.67<\/td>56.12\u00b10.69<\/td><\/tr>
(4)<\/td>15.06<\/strong>\u00b10.58<\/strong> <\/strong><\/strong><\/td>15.89<\/strong>\u00b10.71<\/strong> (AC<\/strong>50<\/sub><\/strong><\/sub><\/strong>)<\/strong><\/strong><\/td><\/tr>
Chloroquine<\/td>1.25\u00b10.17                                          <\/td>1.92\u00b10.19<\/td><\/tr><\/tbody><\/table>
Bile\u015fiklerin glutatyon red\u00fcktaz aktivite IC50<\/sub> de\u011ferleri<\/figcaption><\/figure>\n\n\n\n

Bile\u015fikler (P, 1-3)<\/strong> glutatyon red\u00fcktaz enzimine kar\u015f\u0131 inhibit\u00f6r etkisi g\u00f6sterirken, 4 nolu bile\u015fik enzim aktivat\u00f6r\u00fc olarak davranmaktad\u0131r. \u015eekil 2.2c de (1)<\/strong> ve (4 )<\/strong> nolu Schiff bazlar\u0131na ait ekmek mayas\u0131 GR % aktivite\u2013 deri\u015fim grafi\u011fi verilmektedir. Grafiklerden g\u00f6r\u00fcld\u00fc\u011f\u00fc gibi (1)<\/strong> inhibitor, (4)<\/strong> aktivat\u00f6r olarak davranmaktad\u0131r. En iyi inhibit\u00f6r etkisini (1)<\/strong> nolu Schiff baz\u0131 g\u00f6stermektedir. T\u00fcm bile\u015fikler, ekmek mayas\u0131 GR enzimine kar\u015f\u0131 daha fazla aktivite g\u00f6stermektedir.<\/p>\n<\/div><\/div>\n\n\n\n

\"\"\"\"<\/div><\/figure>\n\n\n\n

Yukar\u0131daki grafikler L1 ligand\u0131n\u0131n ve L4 ligand\u0131n\u0131n birbirinin z\u0131tt\u0131 etki yapt\u0131\u011f\u0131n\u0131 g\u00f6stermektedir. Glutatyon red\u00fcktaz<\/strong> enzim aktivitesi uygulanan ligandlara g\u00f6re aktivat\u00f6r ve inhibit\u00f6r etki g\u00f6stermi\u015ftir. Bu konunun daha kapsaml\u0131 analizinin yap\u0131ld\u0131\u011f\u0131 Antimalaryal \u0130la\u00e7lar \u2013 Yap\u0131 Bazl\u0131 \u0130la\u00e7 Tasar\u0131m\u0131<\/a> yaz\u0131nda bu konu nedenleri ile birlikte tart\u0131\u015f\u0131lmaktad\u0131r.<\/p>\n","protected":false},"excerpt":{"rendered":"

Glutatyon red\u00fcktaz aktivitesi ile ilgili bu yaz\u0131daki ligandlar\u0131n sentezi, yap\u0131 […]<\/p>\n","protected":false},"author":1,"featured_media":52,"comment_status":"open","ping_status":"open","sticky":false,"template":"single-hero-cover.php","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[91],"tags":[208,211,8,209,102,104,92,210,101,125,143,106,105,103,155],"class_list":["post-95","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-enzimler-biyokimya","tag-antioksidan","tag-antiparazit","tag-docking","tag-enzim","tag-enzim-aktivitesi","tag-enzim-inhibisyonu","tag-enzimler","tag-glutatyon","tag-glutatyon-reduktaz","tag-ilac","tag-ilac-gelistirme","tag-ilac-kesfi","tag-molekuler-docking","tag-schiff-bazlari","tag-yanastirma"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=\/wp\/v2\/posts\/95","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=95"}],"version-history":[{"count":2,"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=\/wp\/v2\/posts\/95\/revisions"}],"predecessor-version":[{"id":7364,"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=\/wp\/v2\/posts\/95\/revisions\/7364"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=\/wp\/v2\/media\/52"}],"wp:attachment":[{"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=95"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=95"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.ortaakarsu.net\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=95"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}