Структурные механизмы взаимодействия цианоакрилатов с тубулином растений

Структурные механизмы взаимодействия цианоакрилатов с тубулином растений

Структурные механизмы, обусловливающие специфическое связывание соединений цианоакрилатной природы тубулином высших растений, исследованы на примере взаимодействия этил-(2Z)-3-амино-2-циано-4-этилгекса-2-еноата (СА1) и изопропил-(2Z)-3-амино-2-циано-4-этилгекса-2-еноата (СА2) с α-тубулином Arabidops...

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Datum:2014
Hauptverfasser: Ныпорко, А.Ю., Блюм, Я.Б.
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Veröffentlicht: Інститут клітинної біології та генетичної інженерії НАН України 2014
Schriftenreihe:Цитология и генетика
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Оригинальные работы
Online Zugang:https://nasplib.isofts.kiev.ua/handle/123456789/126613
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Zitieren:Структурные механизмы взаимодействия цианоакрилатов с тубулином растений / А.Ю. Ныпорко, Я.Б. Блюм // Цитология и генетика. — 2014. — Т. 48, № 1. — С. 10-17. — Бібліогр.: 31 назв. — рос.

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Digital Library of Periodicals of National Academy of Sciences of Ukraine
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spelling nasplib_isofts_kiev_ua-123456789-1266132025-02-09T22:49:54Z Структурные механизмы взаимодействия цианоакрилатов с тубулином растений Structural mechanisms of interaction of cyanolcrylates with plant tubulin Ныпорко, А.Ю. Блюм, Я.Б. Оригинальные работы Структурные механизмы, обусловливающие специфическое связывание соединений цианоакрилатной природы тубулином высших растений, исследованы на примере взаимодействия этил-(2Z)-3-амино-2-циано-4-этилгекса-2-еноата (СА1) и изопропил-(2Z)-3-амино-2-циано-4-этилгекса-2-еноата (СА2) с α-тубулином Arabidopsis thaliana. Выявлено, что цианогруппа в составе цианоакрилатов является функциональным аналогом нитрильной группы, определяющей процессы специфического взаимодействия с тубулином растений у динитроанилиновых соединений. На основании данных флуктуации пространственной структуры, динамики водородных связей и энергии взаимодействия СА1 и СА2 выявлен наиболее вероятный тип связывания этих соединений с растительным α-тубулином и охарактеризован соответствующий сайт взаимодействия. При этом 7 из 10 остатков в составе этого сайта (Глн-133, Асн-249, Вал-250, Асп-251, Вал-252, Асн-253 и Глу-254) являются облигатными компоненттами сайта связывания динитроанилинов на поверхности α-тубулина растений. Таким образом, охарактеризованный нами сайт связывания на поверхности α-тубулина способен распознавать и специфически связывать вещества, кардинально отличные по своей химической природе и не имеющие общих фармакофорных групп, при условии определенного сходства их электростатической топологии. The structural mechanisms underlying the specific binding of cyanoacrylate compounds with tubulin of higher plants have been studied by the example of the interaction of ethyl-(2Z)-3-amino-2-cyano-4-ethylhex-2-enoate (CA1) and isopropyl-(2Z)-3-amino-2-cyano-4-ethylhex-2-enoate (CA2) with Arabidopsis thaliana α-tubulin. It was revealed that the cyano group of cyanoacrylates is a functional analogue of the nitrile group, which determines the processes of specific interaction with plant tubulin for dinitroaniline compounds. Based on data on spatial structure fluctuations, the dynamics of hydrogen bonds and the interaction energy of CA1 and CA2 (the most probable binding mode for these compounds with plant α-tubulin) was identified and the appropriate site of interaction was characterized. Seven out of ten residues composing this site (Gln-133, Asn-249, Val-250, Asp-251, Val-252, Asn-253, and Glu-254) are obligatory components of dinitroanilines’ binding site on the plant α-tubulin surface. Thus, the binding site on the α-tubulin surface characterized by us is able to recognize and specifically bind substances, which are cardinally different by their chemical nature and have no common pharmacophore groups, under the condition of a certain similarity of their electrostatic topology. 2014 Article Структурные механизмы взаимодействия цианоакрилатов с тубулином растений / А.Ю. Ныпорко, Я.Б. Блюм // Цитология и генетика. — 2014. — Т. 48, № 1. — С. 10-17. — Бібліогр.: 31 назв. — рос. 0564-3783 DOI: 10.3103/S009545271401006X https://nasplib.isofts.kiev.ua/handle/123456789/126613 577.322.4: 5: 7+576.311.348.7 ru Цитология и генетика application/pdf Інститут клітинної біології та генетичної інженерії НАН України
institution Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection DSpace DC
language Russian
topic Оригинальные работы
Оригинальные работы
spellingShingle Оригинальные работы
Оригинальные работы
Ныпорко, А.Ю.
Блюм, Я.Б.
Структурные механизмы взаимодействия цианоакрилатов с тубулином растений
Цитология и генетика
description Структурные механизмы, обусловливающие специфическое связывание соединений цианоакрилатной природы тубулином высших растений, исследованы на примере взаимодействия этил-(2Z)-3-амино-2-циано-4-этилгекса-2-еноата (СА1) и изопропил-(2Z)-3-амино-2-циано-4-этилгекса-2-еноата (СА2) с α-тубулином Arabidopsis thaliana. Выявлено, что цианогруппа в составе цианоакрилатов является функциональным аналогом нитрильной группы, определяющей процессы специфического взаимодействия с тубулином растений у динитроанилиновых соединений. На основании данных флуктуации пространственной структуры, динамики водородных связей и энергии взаимодействия СА1 и СА2 выявлен наиболее вероятный тип связывания этих соединений с растительным α-тубулином и охарактеризован соответствующий сайт взаимодействия. При этом 7 из 10 остатков в составе этого сайта (Глн-133, Асн-249, Вал-250, Асп-251, Вал-252, Асн-253 и Глу-254) являются облигатными компоненттами сайта связывания динитроанилинов на поверхности α-тубулина растений. Таким образом, охарактеризованный нами сайт связывания на поверхности α-тубулина способен распознавать и специфически связывать вещества, кардинально отличные по своей химической природе и не имеющие общих фармакофорных групп, при условии определенного сходства их электростатической топологии.
format Article
author Ныпорко, А.Ю.
Блюм, Я.Б.
author_facet Ныпорко, А.Ю.
Блюм, Я.Б.
author_sort Ныпорко, А.Ю.
title Структурные механизмы взаимодействия цианоакрилатов с тубулином растений
title_short Структурные механизмы взаимодействия цианоакрилатов с тубулином растений
title_full Структурные механизмы взаимодействия цианоакрилатов с тубулином растений
title_fullStr Структурные механизмы взаимодействия цианоакрилатов с тубулином растений
title_full_unstemmed Структурные механизмы взаимодействия цианоакрилатов с тубулином растений
title_sort структурные механизмы взаимодействия цианоакрилатов с тубулином растений
publisher Інститут клітинної біології та генетичної інженерії НАН України
publishDate 2014
topic_facet Оригинальные работы
url https://nasplib.isofts.kiev.ua/handle/123456789/126613
citation_txt Структурные механизмы взаимодействия цианоакрилатов с тубулином растений / А.Ю. Ныпорко, Я.Б. Блюм // Цитология и генетика. — 2014. — Т. 48, № 1. — С. 10-17. — Бібліогр.: 31 назв. — рос.
series Цитология и генетика
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AT blûmâb strukturnyemehanizmyvzaimodeistviâcianoakrilatovstubulinomrastenii
AT nyporkoaû structuralmechanismsofinteractionofcyanolcrylateswithplanttubulin
AT blûmâb structuralmechanismsofinteractionofcyanolcrylateswithplanttubulin
first_indexed 2025-12-01T13:22:24Z
last_indexed 2025-12-01T13:22:24Z
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fulltext 10 ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 1 Ñòðóêòóðíûå ìåõàíèçìû, îáóñëîâëèâàþùèå ñïåöèôè- ÷åñêîå ñâÿçûâàíèå ñîåäèíåíèé öèàíîàêðèëàòíîé ïðèðî- äû òóáóëèíîì âûñøèõ ðàñòåíèé, èññëåäîâàíû íà ïðè- ìåðå âçàèìîäåéñòâèÿ ýòèë-(2Z)-3-àìèíî-2-öèàíî-4- ýòèëãåêñà-2-åíîàòà (ÑÀ1) è èçîïðîïèë-(2Z)-3-àìèíî- 2-öèàíî-4-ýòèëãåêñà-2-åíîàòà (ÑÀ2) ñ -òóáóëèíîì Arabidopsis thaliana. Âûÿâëåíî, ÷òî öèàíîãðóïïà â ñîñ- òàâå öèàíîàêðèëàòîâ ÿâëÿåòñÿ ôóíêöèîíàëüíûì àíà- ëîãîì íèòðèëüíîé ãðóïïû, îïðåäåëÿþùåé ïðîöåññû ñïå- öèôè÷åñêîãî âçàèìîäåéñòâèÿ ñ òóáóëèíîì ðàñòåíèé ó äèíèòðîàíèëèíîâûõ ñîåäèíåíèé. Íà îñíîâàíèè äàí- íûõ ôëóêòóàöèè ïðîñòðàíñòâåííîé ñòðóêòóðû, äèíà- ìèêè âîäîðîäíûõ ñâÿçåé è ýíåðãèè âçàèìîäåéñòâèÿ ÑÀ1 è ÑÀ2 âûÿâëåí íàèáîëåå âåðîÿòíûé òèï ñâÿçûâà- íèÿ ýòèõ ñîåäèíåíèé ñ ðàñòèòåëüíûì -òóáóëèíîì è îõàðàêòåðèçîâàí ñîîòâåòñòâóþùèé ñàéò âçàèìîäåé- ñòâèÿ. Ïðè ýòîì 7 èç 10 îñòàòêîâ â ñîñòàâå ýòîãî ñàéòà (Ãëí-133, Àñí-249, Âàë-250, Àñï-251, Âàë-252, Àñí-253 è Ãëó-254) ÿâëÿþòñÿ îáëèãàòíûìè êîìïîíåí- òàìè ñàéòà ñâÿçûâàíèÿ äèíèòðîàíèëèíîâ íà ïîâåðõ- íîñòè -òóáóëèíà ðàñòåíèé. Òàêèì îáðàçîì, îõàðàê- òåðèçîâàííûé íàìè ñàéò ñâÿçûâàíèÿ íà ïîâåðõíîñòè -òóáóëèíà ñïîñîáåí ðàñïîçíàâàòü è ñïåöèôè÷åñêè ñâÿçûâàòü âåùåñòâà, êàðäèíàëüíî îòëè÷íûå ïî ñâîåé õèìè÷åñêîé ïðèðîäå è íå èìåþùèå îáùèõ ôàðìàêî- ôîðíûõ ãðóïï, ïðè óñëîâèè îïðåäåëåííîãî ñõîäñòâà èõ ýëåêòðîñòàòè÷åñêîé òîïîëîãèè. Êëþ÷åâûå ñëîâà: òóáóëèí, öèàíîàêðèëàòû, ñïåöèôè- ÷åñêîå âçàèìîäåéñòâèå, ìåõàíèçìû ñâÿçûâàíèÿ, ýëåêòðî- ñòàòè÷åñêàÿ òîïîëîãèÿ. Ââåäåíèå. Èíòåðåñ èññëåäîâàòåëåé ê ãåðáè- öèäíîé àêòèâíîñòè ïðîèçâîäíûõ öèàíîàêðè- ëàòîâ, îòðûòîé áîëåå 30 ëåò íàçàä [1], îñîáåííî óñèëèëñÿ â ïîñëåäíåå äåñÿòèëåòèå. Íà÷èíàÿ ñ 2003 ã., íà îñíîâå ïðîèçâîäíûõ öèàíîàêðèëà- òîâ, ñîäåðæàùèõ èçîêñàçîëüíûå çàìåñòèòåëè, ñîçäàí öåëûé ðÿä ýôôåêòèâíûõ, êîììåð÷åñêè çíà÷èìûõ ãåðáèöèäîâ [2, 3]. Ïîñòóëèðóåòñÿ, ÷òî ãåðáèöèäíàÿ àêòèâíîñòü áîëüøèíñòâà èç íèõ áàçèðóåòñÿ íà ñïîñîáíîñòè èíãèáèðîâàòü òðàíñ- ïîðò ýëåêòðîíîâ â ðåàêöèîííîì öåíòðå ôîòî- ñèñòåìû II [1, 4]. Îäíàêî ó íåêîòîðûõ öèàíîàê- ðèëàòíûõ ñîåäèíåíèé, â ÷àñòíîñòè ó ýòèë-(2Z)- 3-àìèíî-2-öèàíî-4-ýòèëãåêñà-2-åíîàòà (ÑÀ1) è èçîïðîïèë-(2Z)-3-àìèíî-2-öèàíî-4-ýòèëãåê- ñà-2-åíîàòà (ÑÀ2) (ðèñ. 1), âûÿâëåí ñîâåðøåííî èíîé ìåõàíèçì ãåðáèöèäíîãî äåéñòâèÿ: ïîäîá- íî äèíèòðîàíèëèíîâûì è ôîñôîðîàìèäíûì ñî- åäèíåíèÿì ýòè âåùåñòâà îáëàäàþò ñïîñîáíîñ- òüþ ýôôåêòèâíî ðàçðóøàòü ìèêðîòðóáî÷êè ðàñ- òèòåëüíîé êëåòêè [5]. Êàê èçâåñòíî, â ñèëó ôóíäàìåíòàëüíîãî ñâîéñòâà ìîëåêóë òóáóëèíà – îñíîâíîãî áåëêà ìèêðîòðóáî÷åê – ñïåöèôè÷åñ- êèì îáðàçîì ñâÿçûâàòü íèçêîìîëåêóëÿðíûå ñî- åäèíåíèÿ ðàçëè÷íîé ïðèðîäû, ñèñòåìà ìèêðî- òðóáî÷åê ÿâëÿåòñÿ ñâîåîáðàçíûì «óçêèì ìåñòîì» â îòâåòå ýóêàðèîòè÷åñêîé êëåòêè íà âîçäåéñòâèå ìíîãèõ òèïîâ õèìè÷åñêèõ âåùåñòâ [6–8]. Èìåí- íî - èëè -ñóáúåäèíèöû òóáóëèíà ÿâëÿþòñÿ ìèøåíÿìè äëÿ øèðîêîãî ñïåêòðà êîììåð÷åñ- êè çíà÷èìûõ ïðåïàðàòîâ, êîòîðûå èñïîëüçóþò- ñÿ â êà÷åñòâå ôóíãèöèäîâ, ãåðáèöèäîâ, ïðî- òèâîîïóõîëåâûõ, àíòèïðîòîçîéíûõ è ïðîòèâî- ãåëüìèíòíûõ ñðåäñòâ [8–10]. Ñëåäñòâèåì òàêîãî âûñîêîñïåöèôè÷åñêîãî âçàèìîäåéñòâèÿ ÿâëÿ- åòñÿ ëèáî ðàçðóøåíèå, ëèáî íåîáðàòèìàÿ ñòà- áèëèçàöèÿ ìèêðîòðóáî÷åê [6–8, 10, 11]. Óñòîé÷èâîñòü ê àíòèòóáóëèíîâûì âåùåñò- âàì â îáùåì ñëó÷àå îáóñëîâëåíà åäèíè÷íûìè àìèíîêèñëîòíûìè çàìåíàìè [9, 12, 13]. Òàêèå çàìåíû â áåëêå ëîêàëèçîâàíû â íåïîñðåäñòâåí- íîé áëèçîñòè îò ñàéòîâ ñâÿçûâàíèÿ ñîåäèíå- íèé, óñòîé÷èâîñòü ê êîòîðûì îíè âûçûâàþò [13].  ñâÿçè ñ ýòèì íåëüçÿ íå îòìåòèòü òîò ôàêò, ÷òî àìèíîêèñëîòíûå çàìåíû â ïîëîæå- íèè 239 ìîëåêóëû -òóáóëèíà, êîòîðûå îáóñëîâ- ëèâàþò ïîÿâëåíèå áèîòèïîâ, óñòîé÷èâûõ ê äè- íèòðîàíèëèíîâûì ãåðáèöèäàì ó ãóñèíîé òðàâû (Eleusine indica L. Gaerth.) [14, 15] è ùåòèííèêà çåëåíîãî (Setaria viridis L. Beauv.) [16], òàêæå ÿâ- ëÿþòñÿ ïðè÷èíîé óñòîé÷èâîñòè ê ÑÀ1 è ÑÀ2 ó ìîäåëüíîãî ðàñòåíèÿ ðåçóøêè Òàëÿ (Arabidopsis thaliana L.) è óïîìÿíóòîé ãóñèíîé òðàâû [5]. Ñ âûñîêîé âåðîÿòíîñòüþ ìîæíî ïðåäïîëîæèòü, УДК 577.322.4: 5: 7+576.311.348.7 А.Ю. НЫПОРКО 1, Я.Б. БЛЮМ 2 1 Киевский национальный университет имени Тараса Шевченко 2 Институт пищевой биотехнологии и геномики НАН Украины, Киев E-mail: dfnalex@gmail.com СТРУКТУРНЫЕ МЕХАНИЗМЫ ВЗАИМОДЕЙСТВИЯ ЦИАНОАКРИЛАТОВ С ТУБУЛИНОМ РАСТЕНИЙ © À.Þ. ÍÛÏÎÐÊÎ, ß.Á. ÁËÞÌ, 2014 11 Ñòðóêòóðíûå ìåõàíèçìû âçàèìîäåéñòâèÿ öèàíîàêðèëàòîâ ñ òóáóëèíîì ðàñòåíèé ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 1 ÷òî îáà êëàññà ñîåäèíåíèé èìåþò îáùèé ñàéò ñâÿçûâàíèÿ íà ïîâåðõíîñòè ðàñòèòåëüíîãî - òóáóëèíà. Ó÷èòûâàÿ, ÷òî â ñîñòàâå äèíèòðîàíè- ëèíîâ è öèàíîàêðèëàòîâ îòñóòñòâóþò îáùèå ñòðóêòóðíûå êîìïîíåíòû, äåòàëüíîå èññëåäî- âàíèå ñòðóêòóðíûõ ìåõàíèçìîâ âçàèìîäåéñò- âèÿ öèàíîàêðèëàòîâ ñ ìîëåêóëàìè -òóáóëèíà ïðåäñòàâëÿåò íåñîìíåííûé èíòåðåñ, ïðè÷åì êàê ñ òåîðåòè÷åñêîé (óãëóáëåííîå ïîíèìàíèå çàêîíîìåðíîñòåé ôîðìèðîâàíèÿ íà ïîâåðõíîñ- òè áåëêîâ ñàéòîâ ñïåöèôè÷åñêîãî ñâÿçûâàíèÿ íèçêîìîëåêóëÿðíûõ âåùåñòâ), òàê è ñ ïðèê- ëàäíîé (ðàçðàáîòêà ãåðáèöèäîâ ñ ïîâûøåííîé ñåëåêòèâíîñòüþ) òî÷åê çðåíèÿ, ÷òî, ñîáñòâåí- íî, è îïðåäåëèëî êîíå÷íóþ öåëü íàñòîÿùåãî èññëåäîâàíèÿ. Ìàòåðèàëû è ìåòîäû. Ïðîìîäåëè ïðîñòðàí- ñòâåííîé ñòðóêòóðû ñîåäèíåíèé ýòèë-(2Z)-3- àìèíî-2-öèàíî-4-ýòèëãåêñà-2-åíîàòà (ÑÀ1) è èçîïðîïèë-(2Z)-3-àìèíî-2-öèàíî-4-ýòèë-ãåê- ñà-2-åíîàòà (ÑÀ2), à òàêæå ñîîòâåòñòâóþùèå èì ôàéëû òîïîëîãèè áûëè ðåêîíñòðóèðîâàíû ñ ïî- ìîùüþ âåá-èíñòðóìåíòà PRODRG (http://da- vapc1.bioch.dundee.ac.uk/prodrg/) [17]. Ýëåêòðîí- íûå ñâîéñòâà ÑÀ1 è ÑÀ2, â òîì ÷èñëå çíà÷åíèÿ ÷àñòè÷íûõ çàðÿäîâ íà àòîìàõ, ðàññ÷èòàíû íåýì- ïèðè÷åñêèì ìåòîäîì ôóíêöèîíàëà ïëîòíîñòè M062õ [18] ñ èñïîëüçîâàíèåì áàçîâîãî íàáîðà 6-31 + g(d,p). Ïðîñòðàíñòâåííàÿ ñòðóêòóðà öèàíî- àêðèëàòîâ îïòèìèçèðîâàíà ïðè ïîìîùè óïîìÿ- íóòîãî ìåòîäà M062õ/6-31+g(d,p) ñ ïðèìåíåíèåì àëãîðèòìà Áåðíè [19]. Ðàññ÷èòàííûå çíà÷åíèÿ ÷àñòè÷íûõ çàðÿäîâ èñïîëüçîâàíû äëÿ ìîäèôè- êàöèè ñîîòâåòñòâóþùèõ ôàéëîâ òîïîëîãèè. Îïòèìèçèðîâàííûå ñòðóêòóðû ÑÀ1 è ÑÀ2 ïðèìåíÿëè äëÿ ðåêîíñòðóêöèè èõ êîìïëåêñîâ ñ ìîëåêóëîé 1-òóáóëèíà A. thaliana. Ïðîñò- ðàíñòâåííàÿ ñòðóêòóðà ýòîãî áåëêà, èñïîëüçî- âàííàÿ â êà÷åñòâå ìèøåíè ñâÿçûâàíèÿ öèàíî- àêðèëàòîâ, ðåêîíñòðóèðîâàíà è îïèñàíà â íà- øåé ïðåäûäóùåé ðàáîòå [20]. Ïîñëåäîâàòåëü- íîñòü 1-òóáóëèíà A. thaliana ñîäåðæèò 450 àìè- íîêèñëîòíûõ îñòàòêîâ, äåïîíèðîâàíà â áàí- êå àìèíîêèñëîòíûõ ïîñëåäîâàòåëüíîñòåé Uniprot (www.uniprot.org), íîìåð äîñòóïà P11139. Èñõîäíîå ïîçèöèîíèðîâàíèå ëèãàíäîâ â ñàé- òå âçàèìîäåéñòâèÿ íà ïîâåðõíîñòè -òóáóëè- íà îñóùåñòâëÿëè ñ ïîìîùüþ ìåòîäà ìóëüòè- êîïèéíîãî îäíîâðåìåííîãî ïîèñêà [21]. Îïòè- ìèçàöèþ ãåîìåòðèè êîìïëåêñîâ ÑÀ1 è ÑÀ2 ñ -òóáóëèíîì ïðîèçâîäèëè ïóòåì ìèíèìèçàöèè ïîòåíöèàëüíîé ýíåðãèè ïðè ïîìîùè ìåòîäà L-BFGS [22] ïîñðåäñòâîì óïîòðåáëåíèÿ ìîäóëÿ mdrun ïðîãðàììíîãî îáåñïå÷åíèÿ GROMACS 4.0.3 [23]. Äëÿ ýòîãî èñïîëüçîâàëè ñèëîâîå ïî- ëå GROMACS, ìàêñèìàëüíîå êîëè÷åñòâî øà- ãîâ îïòèìèçàöèè ñîñòàâëÿëî 1000. Êðèòåðèÿìè ñõîäèìîñòè âûñòóïàëè ïðèíÿòûå ïî óìîë÷à- íèþ çíà÷åíèÿ ìàêñèìàëüíîé ñèëû íà àòîìàõ emtol = 10 êÄæ·ìîëü–1·íì–1 è ìàêñèìàëüíîé äëèíû øàãà 0,01 íì. Âñå ðàñ÷åòû ïðîâîäèëè â ýêñïëèöèòíîì (ðåàëüíîì) âîäíîì îêðóæåíèè. Äëÿ ýòîãî âîêðóã êîìïëåêñîâ öèàíîàêðèëàòîâ ñ -òóáóëèíîì ãåíåðèðîâàëè ïåðèîäè÷åñêèå áîê- ñû ðàçìåðîì 7.25700 × 7.13400 × 7.91200 íì3 (ìîäóëü editconf), êîòîðûå çàïîëíÿëèñü ìîëåêó- ëàìè âîäû (ìîäóëü genbox ïðîãðàììíîãî îáåñ- ïå÷åíèÿ GROMACS). Ìàêñèìàëüíûé ðàçìåð èñ- ñëåäóåìîé ñèñòåìû ñîñòàâëÿë 38 374 àòîìà. Îïòèìèçèðîâàííûå êîìïëåêñû âîâëåêàëèñü â ðàñ÷åòû îãðàíè÷åííîé (position restrained) ìî- ëåêóëÿðíîé äèíàìèêè â èíòåðâàëå 20 ïñ è íå- îãðàíè÷åííîé ìîëåêóëÿðíîé äèíàìèêè (ÌÄ) â èíòåðâàëå 12 íñ ïðè òåìïåðàòóðå 300 Ê. Èñ- õîäíûå ñêîðîñòè ãåíåðèðîâàëè ñîãëàñíî ðàñ- ïðåäåëåíèþ Ìàêñâåëëà-Áîëüöìàíà ïðè òåìïå- ðàòóðå 300 K. Äëÿ èíòåãðèðîâàíèÿ óðàâíåíèé äâèæåíèÿ èñïîëüçîâàëè êëàññè÷åñêèé leap-frog àëãîðèòì [24], äëÿ ó÷åòà ýëåêòðîñòàòè÷åñêèõ âçàèìîäåéñòâèé – ìåòîä ñåòè ÷àñòèö Ýâàëü- Ðèñ. 1. Ñòðóêòóðû öèàíîàêðèëàòîâ ÑÀ1 (à) è ÑÀ2 (á) 12 À.Þ. Íûïîðêî, ß.Á. Áëþì ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 1 äà [25], äëÿ ó÷åòà Âàí-äåð-âààëüñîâûõ âçàèìî- äåéñòâèé – ìåòîä ðàäèóñîâ îòñå÷åíèÿ. Òåìïå- ðàòóðó â ñèñòåìå ñòàáèëèçèðîâàëè ñ ïîìîùüþ òåðìîñòàòà ìàñøòàáèðîâàíèÿ ñêîðîñòåé [26]. Ñîîáùåíèÿ îá îøèáêàõ è íåäîïóñòèìûõ ñòå- ðè÷åñêèõ ïàðàìåòðàõ èññëåäóåìûõ ñèñòåì íà ïðîòÿæåíèè ïðîöåäóð ðàñ÷åòà îòñóòñòâîâàëè. Äëÿ ïîñëåäóþùåé êîëè÷åñòâåííîé îöåíêè ýíåðãèè ñâÿçûâàíèÿ ðàññ÷èòàëè ìîëåêóëÿðíóþ äèíàìèêó öèàíîàêðèëàòíûõ ñîåäèíåíèé â ñîëü- âàòèðîâàííîì ñîñòîÿíèè. Ïðîöåäóðà ðàñ÷åòà ìîëåêóëÿðíîé äèíàìèêè ñîëüâàòèðîâàííûõ öèà- íîàêðèëàòîâ áûëà òàêîé æå, êàê ïðè ðàñ÷åòå êîìïëåêñîâ «òóáóëèí – öèàíîàêðèëàò», çà èñê- ëþ÷åíèåì ðàçìåðà ïåðèîäè÷åñêîãî áîêñà, â êî- òîðûé ïîìåùàëè èññëåäóåìûå ñîåäèíåíèÿ – 2.52500 × 2.72800 × 2.44900 íì3 äëÿ ÑÀ1 è 2.60200 × 2.79000 × 2.58100 íì3 äëÿ ÑÀ2. Ôëóêòóàöèè ñòðóêòóðû ñîåäèíåíèé â ñîñòà- âå êîìïëåêñîâ îöåíèâàëè ïðè ïîìîùè ìåòî- äà rho [27] c èñïîëüçîâàíèåì ìîäóëÿ g_rms ïðîãðàììíîãî îáåñïå÷åíèÿ GROMACS. Ýíåð- ãèè âçàèìîäåéñòâèÿ ðàññ÷èòûâàëè â âèäå ðàç- íîñòè íåâàëåíòíûõ âêëàäîâ â ïîòåíöèàëüíóþ ýíåðãèþ öèàíîàêðèëàòîâ â ñâîáîäíîì (ñîëü- âàòèðîâàííîì) è ñâÿçàííîì ñ áåëêîì ñîñòîÿ- íèè (ìîäóëü g_energy ïðîãðàììíîãî îáåñïå÷å- íèÿ GROMACS). Äèíàìèêó âîäîðîäíûõ ñâÿ- çåé ðàññ÷èòûâàëè ñ ïîìîùüþ ìîäóëÿ g_hbond. Âèçóàëèçàöèþ ñòðóêòóðíûõ äàííûõ îñóùåñò- âëÿëè ñ ïðèìåíåíèåì ïðîãðàììíîãî îáåñïå- ÷åíèÿ Accelrys DS Vizualiser (âåðñèè 2.0 è 3.5) (www.accelrys.com). Ðåçóëüòàòû èññëåäîâàíèé è èõ îáñóæäåíèå.  ñòðóêòóðå èññëåäîâàííûõ ìîëåêóë öèàíî- àêðèëàòîâ ñîäåðæàòñÿ òðè êîìïîíåíòà, îáëà- äàþùèõ âûðàæåííûìè îòðèöàòåëüíûìè çíà÷å- íèÿìè ýëåêòðîñòàòè÷åñêîãî ïîòåíöèàëà. Ê íèì îòíîñÿòñÿ àòîì àçîòà â ñîñòàâå öèàíîãðóïïû, àòîì êèñëîðîäà ïðè äâîéíîé ñâÿçè ñëîæíî- ýôèðíîé ãðóïïû è àòîì àçîòà â ñîñòàâå àìè- íîãðóïïû (ðèñ. 2, ñì. âêëåéêó). Ïîòåíöèàëüíàÿ âàæíîñòü ýòèõ êîìïîíåíòîâ äëÿ ñïåöèôè÷åñ- êîãî âçàèìîäåéñòâèÿ ñ ìîëåêóëîé ðàñòèòåëüíîãî -òóáóëèíà îáóñëîâëåíà ñëåäóþùèìè ñîîáðà- æåíèÿìè. Ðàíåå íàìè óñòàíîâëåíî, ÷òî êëþ÷å- âîé ñîñòàâëÿþùåé ñàéòà ñâÿçûâàíèÿ äèíèòðî- àíèëèíîâ íà ïîâåðõíîñòè -òóáóëèíà ÿâëÿåò- ñÿ ïîëîæèòåëüíî çàðÿæåííàÿ ïîëîñòü, ðàñïî- ëîæåííàÿ â îáëàñòè èíòåðäèìåðíîãî êîíòàêòà ìåæäó ñóáúåäèíèöàìè òóáóëèíà [28–30]. Ýòà îáëàñòü ñëóæèò ìåñòîì ñâÿçûâàíèÿ íèòðèëüíîé ãðóïïû â ñîñòàâå äèíèòðîàíèëèíîâûõ (à òàê- æå ôîñôîðîàìèäíûõ) ñîåäèíåíèé, êîòîðàÿ â ñâîþ î÷åðåäü õàðàêòåðèçóåòñÿ ÿðêî âûðàæåí- íûìè îòðèöàòåëüíûìè çíà÷åíèÿìè ÷àñòè÷íûõ çàðÿäîâ íà àòîìàõ êèñëîðîäà è, êàê ñëåäñò- âèå, îòðèöàòåëüíûì ýëåêòðîñòàòè÷åñêèì ïîòåí- öèàëîì â öåëîì. Òàêèì îáðàçîì, åñëè äè- íèòðîàíèëèíû è öèàíîàêðèëàòû èìåþò îáùèé ñàéò ñâÿçûâàíèÿ íà ïîâåðõíîñòè ìîëåêóëû òó- áóëèíà, òî êðèòè÷åñêóþ ðîëü â ïðîöåññå ñâÿ- çûâàíèÿ äîëæåí èãðàòü èìåííî îòðèöàòåëüíî çàðÿæåííûé ñòðóêòóðíûé êîìïîíåíò â ñîñòàâå ìîëåêóëû öèàíîàêðèëàòà [28–30]. Íèòðèëüíûõ ãðóïï â ñîñòàâå èññëåäóåìûõ ïðåäñòàâèòåëåé öèàíîàêðèëàòîâ íå èìååòñÿ. Èç òðåõ îòðèöàòåëüíî çàðÿæåííûõ êîìïî- íåíòîâ â ñîñòàâå èññëåäóåìûõ öèàíîàêðèëàòîâ àìèíîãðóïïà ïðåäñòàâëÿåòñÿ ìàëîâåðîÿòíûì êàíäèäàòîì íà ðîëü êëþ÷åâîãî êîìïîíåíòà ðàñ- ïîçíàâàíèÿ â ñèëó òîãî, ÷òî îòðèöàòåëüíî çà- ðÿæåííûé àòîì àçîòà â ñîñòàâå ýòîé ãðóïïû ýêðàíèðóåòñÿ àòîìàìè âîäîðîäà, êîòîðûå íåñóò ïîëîæèòåëüíûå çàðÿäû.  ðåçóëüòàòå îáùèé ýëåêòðîñòàòè÷åñêèé ïîòåíöèàë ãðóïïû îêàçû- âàåòñÿ ïîëîæèòåëüíûì. Cîãëàñíî ðåçóëüòàòàì ìóëüòèêîïèéíîãî ïîçèöèîíèðîâàíèÿ ëèãàíäîâ â ñàéòå âçàèìîäåéñòâèÿ àòîì êèñëîðîäà ïðè äâîéíîé ñâÿçè ñëîæíîýôèðíîé ãðóïïû âî âñåõ ñëó÷àÿõ îêàçûâàåòñÿ îðèåíòèðîâàííûì íàðóæó, à íå âíóòðü ïîëîñòè âçàèìîäåéñòâèÿ. Ñëåäîâà- òåëüíî, îí òîæå íå ìîæåò ÿâëÿòüñÿ ðåøàþùèì ôàêòîðîì â ïðîöåññàõ âçàèìîäåéñòâèÿ öèàíî- àêðèëàòîâ ñ -òóáóëèíîì. Òàêèì îáðàçîì, íàèáîëåå âåðîÿòíûì ñòðóê- òóðíûì êîìïîíåíòîì öèàíîàêðèëàòîâ, îïðåäå- ëÿþùèì õàðàêòåð âçàèìîäåéñòâèÿ ýòèõ ñîåäè- íåíèé ñ ìîëåêóëîé -òóáóëèíà ðàñòåíèé, ÿâëÿ- åòñÿ îòðèöàòåëüíî çàðÿæåííûé àçîò öèàíîãðóï- ïû, êîòîðàÿ â äàííîì ñëó÷àå âûñòóïàåò ôóíê- öèîíàëüíûì àíàëîãîì íèòðèëüíîé ãðóïïû, îïðåäåëÿþùåé ïðîöåññû ñïåöèôè÷åñêîãî âçà- èìîäåéñòâèÿ ñ òóáóëèíîì ðàñòåíèé ó äèíèòðî- àíèëèíîâûõ, ðàâíî êàê è ôîñôîðîàìèäíûõ ñîå- äèíåíèé [28–30]. Ðåçóëüòàòû ïîçèöèîíèðîâàíèÿ è ïîñëåäó- þùåãî ðàñ÷åòà ìîëåêóëÿðíîé äèíàìèêè öèàíî- àêðèëàòîâ â ñàéòå âçàèìîäåéñòâèÿ ïîçâîëèëè âûÿâèòü äâå àëüòåðíàòèâíûå îðèåíòàöèè ëèãàí- 13 Ñòðóêòóðíûå ìåõàíèçìû âçàèìîäåéñòâèÿ öèàíîàêðèëàòîâ ñ òóáóëèíîì ðàñòåíèé ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 1 äîâ, êîòîðûå ñîîòâåòñòâóþò äâóì âîçìîæíûì òèïàì ñâÿçûâàíèÿ ýòèõ ñîåäèíåíèé ìîëåêóëîé -òóáóëèíà.  ïåðâîì âàðèàíòå öèàíîãðóïïà îðèåíòèðîâàíà âíóòðü ñàéòà ñâÿçûâàíèÿ è âîâ- ëå÷åíà â ôîðìèðîâàíèå âîäîðîäíûõ ñâÿçåé ñ íåñêîëüêèìè àìèíîêèñëîòíûìè îñòàòêàìè, ðàñïîëîæåííûìè â ãëóáèíå ñàéòà (ðèñ. 3). Âî âòîðîì âàðèàíòå öèàíîãðóïïà ðàñïîëîæåíà ïðàêòè÷åñêè ïàðàëëåëüíî ìîëåêóëÿðíîé ïî- âåðõíîñòè áåëêà è ôîðìèðóåò âîäîðîäíûå ñâÿ- çè ñ îñòàòêàìè, ýêñïîíèðîâàííûìè íà ïî- âåðõíîñòè, â ÷àñòíîñòè ñ Àðã-2 â ñëó÷àå ÑÀ1 (ðèñ. 4). Ñàéò ñâÿçûâàíèÿ ÑÀ1 ïðè âçàèìîäåéñòâèè ïî òèïó ² ñîñòîèò èç àìèíîêèñëîòíûõ îñòàòêîâ Ãëí-133, Àñí-249, Âàë-250, Àñï-251, Âàë-252, Àñí- 253, Ãëó-254, Ãëí-256, Àñí-258, Ëåé-259. Ñåìü èç íèõ, â ÷àñòíîñòè Ãëí-133, Àñí-249, Àñï-251, Àñí-253, Ãëó-254, Ãëí-256 è Àñí-258, ÿâëÿþò- ñÿ îñòàòêàìè, êîòîðûå ïðèíèìàþò íåïîñðåä- ñòâåííîå ó÷àñòèå â ôîðìèðîâàíèè ïðîäîëüíîãî êîíòàêòíîãî èíòåðôåéñà ñ -ñóáúåäèíèöåé ñî- ñåäíåãî äèìåðà [31]. Àòîì àçîòà öèàíîãðóïïû â ñîñòàâå ÑÀ1 ó÷àñòâóåò â ôîðìèðîâàíèè ÷åòû- ðåõ âîäîðîäíûõ ñâÿçåé – ñ âîäîðîäàìè àìèä- íûõ ãðóïï àìèíîêèñëîò Âàë-252, Àñí-253 è Ãëó-254, à òàêæå ñ âîäîðîäîì áîêîâîé àìèíî- ãðóïïû Àñí-253. Êðîìå òîãî, äâå âîäîðîäíûå ñâÿçè ôîðìèðóþòñÿ ñ ó÷àñòèåì âîäîðîäîâ â ñîñòàâå àìèíîãðóïïû öèàíîàêðèëàòà – ñ êèñ- ëîðîäîì êàðáîêñèëüíîé ãðóïïû Ãëó-254 è êèñëîðîäîì êàðáîíèëüíîé ãðóïïû Ãëí-256. Ñàéò ñâÿçûâàíèÿ ÑÀ2 ïðè âçàèìîäåéñòâèè ïî òèïó I çà íåáîëüøèìè èñêëþ÷åíèÿìè ñõîæ ïî àìèíîêèñëîòíîìó ñîñòàâó ñ àíàëîãè÷íûì ñàéòîì ÑÀ1.  ÷àñòíîñòè, â åãî ñîñòàâå îò- ñóòñòâóþò äâà îñòàòêà ãëþòàìèíà (â ïîëîæå- íèÿõ 133 è 256) è ïðèñóòñòâóþò îñòàòêè ëåé- öèíà è ôåíèëàëàíèíà â ïîëîæåíèÿõ 248 è 256 ñîîòâåòñòâåííî. Äâà ïîñëåäíèõ îñòàòêà ïðè- íèìàþò ó÷àñòèå â ôîðìèðîâàíèè ïîâåðõíîñòè ïðîäîëüíîãî êîíòàêòà ìåæäó ñóáúåäèíèöàìè òóáóëèíà [31]. Àçîò öèàíîãðóïïû ÑÀ2 òàêæå ôîðìèðóåò ÷åòûðå âîäîðîäíûå ñâÿçè, ñîâïàäà- þùèå ñî ñâÿçÿìè ó ÑÀ1, îäíàêî âîäîðîäíûå àòîìû àìèíîãðóïïû íå âîâëå÷åíû â îáðàçîâà- íèå âîäîðîäíûõ ñâÿçåé. Ñëåäóåò îòìåòèòü, ÷òî ðÿä îñòàòêîâ, ïðèíèìàþùèõ ó÷àñòèå â ñâÿçû- âàíèè öèàíîàêðèëàòîâ ïî òèïó ², à èìåííî Ãëí-133, Àñí-249, Âàë-250, Àñï-251, Âàë-252, Àñí-253 è Ãëó-254, ÿâëÿþòñÿ îáëèãàòíûìè êîì- ïîíåíòàìè ñàéòà ñâÿçûâàíèÿ äèíèòðîàíèëèíîâ ìîëåêóëîé -òóáóëèíà ðàñòåíèé [30]. Ïðè âçàèìîäåéñòâèè ïî òèïó ²² ñàéò ñâÿ- çûâàíèÿ ÑÀ1 ñôîðìèðîâàí îñòàòêàìè Àðã-2, Ãëí-133, Ëèç-164, Ñåð-165, Àñï-251, Àñí-253 è Ãëó-254 (ðèñ. 4). Ìåíüøåå êîëè÷åñòâî àìèíî- êèñëîò â ýòîì ñëó÷àå ñâÿçàíî ñ îñîáåííîñòÿìè îðèåíòàöèè ëèãàíäà â ñàéòå âçàèìîäåéñòâèÿ è ñóùåñòâåííî åãî ìåíüøåé ïîãðóæåííîñòüþ âíóòðü áåëêîâîé ìîëåêóëû ïî ñðàâíåíèþ ñ âçàèìîäåéñòâèåì ïî òèïó ². Àçîò öèàíîãðóï- ïû ôîðìèðóåò òðè âîäîðîäíûå ñâÿçè: äâå – ñ âîäîðîäàìè áîêîâîé àìèíîãðóïïû Àðã-2 è îäíó – ñ âîäîðîäîì áîêîâîé àìèäíîé ãðóïïû Ãëí-133. Êðîìå òîãî, âîäîðîäû àìèíîãðóïïû ÑÀ1 âîâëå÷åíû â ôîðìèðîâàíèå âîäîðîäíûõ ñâÿçåé ñ êèñëîðîäàìè êàðáîêñèëüíûõ ãðóïï Àñï-251 è Ãëó-254 (ðèñ. 4). Ñàéò ñâÿçûâàíèÿ ÑÀ2 ïðè âçàèìîäåéñòâèè ïî òèïó ²² ðåçêî îò- ëè÷àåòñÿ ïî ñâîåìó ñîñòàâó êàê îò ñàéòà ñâÿ- çûâàíèÿ ÑÀ2 ïåðâîãî òèïà, òàê è îò ñàéòà ñâÿ- çûâàíèÿ ÑÀ1 âòîðîãî òèïà. Îí ñîñòîèò èç àìèíîêèñëîòíûõ îñòàòêîâ Ãëè-246, Àëà-247, Ëåé-248, Àñí-249, Àñí-258, Ëèç-352, Öèñ-353, Ãëè-354, Èëå-355, Òèð-357 (ðèñ. 4). Òàêèì îáðàçîì, èç àìèíîêèñëîòíûõ îñòàòêîâ, âñòðå÷à- þùèõñÿ â ñîñòàâå äðóãèõ ñàéòîâ ñâÿçûâàíèÿ (êàê öèàíîàêðèëàòîâ, òàê è äèíèòðîàíèëèíîâ), â ñîñòàâ ýòîãî ñàéòà âõîäèò òîëüêî îäèí – àñ- ïàðàãèí â ïîëîæåíèè 249. Ïåðåìåùåíèå ëè- ãàíäà â íîâîå ïîëîæåíèå ïðîèñõîäèò äîñòà- òî÷íî ïëàâíî, òåíäåíöèÿ ê îòäåëåíèþ îò áåëêà íà ïðîòÿæåíèè 12 íñ èíòåðâàëà âðåìåíè íå íàáëþäàåòñÿ (÷òî, âïðî÷åì, ñïðàâåäëèâî è äëÿ îñòàëüíûõ òðåõ ñëó÷àåâ). Àçîò öèàíîãðóïïû ôîð- ìèðóåò òðè âîäîðîäíûå ñâÿçè: äâå – ñ âîäîðî- äàìè îñíîâíîé è áîêîâîé àìèäíîé ãðóïïû Àñí-249 è îäíó – ñ âîäîðîäîì áîêîâîé àìèíî- ãðóïïû Ëèç-352. Êðîìå òîãî, îäíà âîäîðîäíàÿ ñâÿçü ôîðìèðóåòñÿ ñ ó÷àñòèåì âîäîðîäà àìèíî- ãðóïïû ÑÀ2 è êèñëîðîäà îñíîâíîé öåïè îñ- òàòêà Öèñ-353. Èç àìèíîêèñëîò êîíòàêòíîãî èíòåðôåéñà â ñàéòàõ ñâÿçûâàíèÿ òèïà II ïðè- ñóòñòâóþò Ãëí-133, Àñï-251, Àñí-253 è Ãëó-254 â ñëó÷àå ÑÀ1 è Àëà-247, Ëåé-248, Àñí-249 è Àñí-258 â ñëó÷àå ÑÀ2. Ôëóêòóàöèè ñòðóêòóðû öèàíîàêðèëàòîâ ïðè ñâÿçûâàíèè ïî òèïó I ïðîÿâëÿþò ÷åòêóþ òåí- äåíöèþ ê ñòàáèëèçàöèè íà îïðåäåëåííîì óðîâ- 14 À.Þ. Íûïîðêî, ß.Á. Áëþì ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 1 íå, ÷òî ñâèäåòåëüñòâóåò î äîñòàòî÷íîé óñòîé- ÷èâîñòè òàêîãî ñâÿçàííîãî ñîñòîÿíèÿ. Ñòàáè- ëèçàöèÿ ôëóêòóàöèé ïðîèñõîäèò â ðàéîíå 5-é íàíîñåêóíäû â ñëó÷àå ÑÀ1 (ïóòåì äâóõñòóïåí- ÷àòîãî ïåðåõîäà, ðèñ. 5, à) è â ðàéîíå 4-é íàíî- ñåêóíäû â ñëó÷àå ÑÀ2 (ðèñ. 5, á). Íàïðîòèâ, ôëóêòóàöèè ñòðóêòóðû öèàíîàêðèëàòîâ, ñâÿçàí- íûõ ïî òèïó II, íå îáíàðóæèâàþò ÷åòêîé òåí- äåíöèè ê êàêîé áû òî íè áûëî ñòàáèëèçàöèè. Îíè ñêîðåå ãîâîðÿò î ïîñëåäîâàòåëüíûõ ïåðå- õîäàõ îò îäíîé êîíôîðìàöèè ëèãàíäà ê äðóãîé, ÷òî ìîæåò ñâèäåòåëüñòâîâàòü î íåóñòîé÷èâîñòè òàêèõ ñîñòîÿíèé. Íèêàêèå âèäèìûå êîððåëÿ- öèè ìåæäó ïîâåäåíèåì ñîåäèíåíèé ÑÀ1 (ðèñ. 5, â) è ÑÀ2 (ðèñ. 5, ã) ïðè ñâÿçûâàíèè òèïà II íå íàáëþäàþòñÿ, ÷òî ïðåäñòàâëÿåòñÿ âïîëíå çà- êîíîìåðíûì ñ ó÷åòîì òîãî, ÷òî â ýòîì ñëó÷àå èññëåäóåìûå ñîåäèíåíèÿ ôàêòè÷åñêè èìåþò ðàçíûå ñàéòû ñâÿçûâàíèÿ, ïåðåêðûâàÿñü ëèøü ïî îäíîé àìèíîêèñëîòå. Îòìåòèì òàêæå, ÷òî ñðåäíèå çíà÷åíèÿ ñðåäíåêâàäðàòè÷åñêèõ îòêëî- íåíèé ïî ðàçíûì òðàåêòîðèÿì íàõîäÿòñÿ â ïðå- äåëàõ îò 1,3 äî 2 Å, ÷òî â ñâîþ î÷åðåäü ñâèäå- òåëüñòâóåò î êîððåêòíîñòè èñõîäíîé ãåîìåòðèè êîìïëåêñîâ «òóáóëèí – öèàíîàêðèëàò». Èññëåäîâàííûå êîìïëåêñû äåìîíñòðèðóþò òàêæå ðàçëè÷íóþ äèíàìèêó ôîðìèðîâàíèÿ/ðàç- ðóøåíèÿ âîäîðîäíûõ ñâÿçåé ìåæäó ìîëåêóëîé -òóáóëèíà è ñâÿçàííûìè ìîëåêóëàìè öèàíî- àêðèëàòîâ.  ñëó÷àå âçàèìîäåéñòâèÿ ïî ïåðâî- ìó òèïó íà äèàãðàììàõ èçìåíåíèÿ êîëè÷åñòâà âîäîðîäíûõ ñâÿçåé ÷åòêî ëîêàëèçîâàíû ñðåä- íèå èõ çíà÷åíèÿ – ïîðÿäêà 5 äëÿ ÑÀ1 (ðèñ. 6, à) è 4 äëÿ ÑÀ2 (ðèñ. 6, á), ÷òî ÿâëÿåòñÿ äîïîë- íèòåëüíûì ñâèäåòåëüñòâîì â ïîëüçó óñòîé÷è- âîñòè ýòèõ êîìïëåêñîâ âî âðåìåíè. Íàïðîòèâ, äèàãðàììû äëÿ êîìïëåêñîâ âòîðîãî òèïà äîñ- òàòî÷íî ðàçìûòû, êîëè÷åñòâî âîäîðîäíûõ ñâÿ- çåé èçìåíÿåòñÿ â âåñüìà øèðîêèõ ïðåäåëàõ – îò 1 äî 7 â ñëó÷àå ÑÀ1 (ðèñ. 6, â) è îò 0 (!) äî 7 â ñëó÷àå ÑÀ2 (ðèñ. 6, ã), ÷òî ïîäòâåðæäàåò èçëîæåííûé òåçèñ î íåóñòîé÷èâîñòè ïðîñòðàí- ñòâåííîé ñòðóêòóðû öèàíîàêðèëàòîâ â êîìï- ëåêñàõ âòîðîãî òèïà è âîçìîæíîé íåñòàáèëü- íîñòè ñàìèõ êîìïëåêñîâ. Íàèáîëåå ñóùåñòâåííî îòëè÷èÿ äâóõ òèïîâ âçàèìîäåéñòâèÿ öèàíîàêðèëàòîâ ñ ìîëåêóëîé -òóáóëèíà ïðîÿâëÿþòñÿ â õàðàêòåðå èçìåíå- íèÿ ïîòåíöèàëüíîé ýíåðãèè ëèãàíäîâ, êîòîðîå ïðîèñõîäèò â ðåçóëüòàòå ñâÿçûâàíèÿ.  ñëó- ÷àå ñâÿçûâàíèÿ ïî òèïó I ïðîèñõîäèò âåñüìà ñóùåñòâåííîå ñíèæåíèå ñðåäíèõ çíà÷åíèé ïî- òåíöèàëüíîé ýíåðãèè îáîèõ ñîåäèíåíèé – íà 38,24 êêàë/ìîëü äëÿ ÑÀ1 è íà 43,34 êêàë/ìîëü äëÿ ÑÀ2. Òàêîé ñäâèã ÿâëÿåòñÿ íàäåæíûì ñâè- Ðèñ. 5. Ôëóêòóàöèè ïðîñòðàíñòâåííîé ñòðóêòóðû öèàíîàêðèëàòíûõ ñîåäèíåíèé â êîìïëåêñå ñ -òó- áóëèíîì A. thaliana: à, á – òèï ñâÿçûâàíèÿ I; â, ã – òèï ñâÿçûâàíèÿ II; ïî âåðòèêàëè – ñðåäíåêâàäðàòè- ÷åñêèå îòêëîíåíèÿ, íì; ïî ãîðèçîíòàëè – âðåìÿ, íñ 15 Ñòðóêòóðíûå ìåõàíèçìû âçàèìîäåéñòâèÿ öèàíîàêðèëàòîâ ñ òóáóëèíîì ðàñòåíèé ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 1 äåòåëüñòâîì ñòàáèëüíîãî êîìïëåêñîîáðàçîâàíèÿ. Ñíèæåíèå ïîòåíöèàëüíîé ýíåðãèè ïðè ñâÿçû- âàíèè ÑÀ1 ïî òèïó II ÿâëÿåòñÿ íåñêîëüêî ìåíü- øèì – 18,9 êêàë/ìîëü, ÷òî â îáùåì ñëó÷àå òî- æå ìîæåò áûòü äîñòàòî÷íûì äëÿ ôîðìèðîâàíèÿ êîìïëåêñà ìåæäó ãåðáèöèäîì è áåëêîì. Îäíàêî ïðè ñâÿçûâàíèè ïî âòîðîìó òèïó ñîåäèíåíèÿ ÑÀ2 åãî ýíåðãèÿ óìåíüøàåòñÿ âñåãî íà 2,3 êêàë/ìîëü, ÷òî ÿâíî íåäîñòàòî÷íî äëÿ îáðàçîâàíèÿ ñêîëü- íèáóäü óñòîé÷èâîãî âî âðåìåíè êîìïëåêñà. Òàêèì îáðàçîì, ìîæíî çàêëþ÷èòü, ÷òî óíè- âåðñàëüíûì ìåõàíèçìîì âçàèìîäåéñòâèÿ ãåð- áèöèäîâ öèàíîàêðèëàòíîé ïðèðîäû ñ ðàñòè- òåëüíûì -òóáóëèíîì c âûñîêîé äîëåé âåðîÿò- íîñòè ÿâëÿåòñÿ ñâÿçûâàíèå ïî òèïó I (ðèñ. 7).  ïîëüçó ýòîãî óòâåðæäåíèÿ ñâèäåòåëüñòâó- åò, âî-ïåðâûõ, ñõîäíûé ñîñòàâ àìèíîêèñëîòíî- ãî îêðóæåíèÿ ÑÀ1 è ÑÀ2 ïðè ñâÿçûâàíèè ïî òèïó I. Ìàëîâåðîÿòíî, ÷òî ñîåäèíåíèÿ, ðàçëè- ÷àþùèåñÿ èñêëþ÷èòåëüíî íàëè÷èåì/îòñóòñòâè- åì îäíîé ìåòèëüíîé ãðóïïû, êîòîðàÿ â äàííîì ñëó÷àå íå ÿâëÿåòñÿ îïðåäåëÿþùåé äëÿ ñïåöè- ôè÷åñêîãî ìåæìîëåêóëÿðíîãî ðàñïîçíàâàíèÿ, áóäóò èìåòü êàðäèíàëüíûå îòëè÷èÿ ñàéòîâ ñâÿ- çûâàíèÿ íà ïîâåðõíîñòè ìîëåêóëû áåëêà. Âî- âòîðûõ, ñòðóêòóðà ÑÀ1 è ÑÀ2 ïðè ñâÿçûâàíèè ïî òèïó I äîñòàòî÷íî ñòàáèëüíà âî âðåìåíè â ïðîòèâîïîëîæíîñòü çíà÷èòåëüíûì ïåðåñòðîé- êàì ïðè âçàèìîäåéñòâèè ïî òèïó II, î ÷åì ñâèäåòåëüñòâóåò ñîîòâåòñòâóþùàÿ ðàçíèöà â äè- íàìêå ÑÊÎ è âîäîðîäíûõ ñâÿçåé ìåæäó ëèãàí- äàìè è áåëêîì. Íî åäâà ëè íå ñàìûì âàæíûì ÿâëÿåòñÿ òîò ôàêò, ÷òî ÑÀ1 è ÑÀ2 ïðè âçàè- ìîäåéñòâèè ïî òèïó I èìåþò áëèçêèå ïî çíà- Ðèñ. 6. Èçìåíåíèå êîëè÷åñòâà âîäîðîäíûõ ñâÿçåé (ïî âåðòèêàëè) ìåæäó öèàíîàêðèëàòíûìè ñîåäèíåíèÿìè è ìîëåêóëîé -òóáóëèíà A. thaliana â ïðîöåññå ìîëåêóëÿðíîé äèíàìèêè ñîîòâåòñòâóþùèõ êîìïëåêñîâ: à, á – òèï ñâÿçûâàíèÿ I; â, ã – òèï ñâÿçûâàíèÿ II; ïî ãîðèçîíòàëè – âðåìÿ, íñ 16 À.Þ. Íûïîðêî, ß.Á. Áëþì ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 1 ÷åíèþ âåëè÷èíû ýíåðãèé ñâÿçûâàíèÿ, òîãäà êàê ïðè âçàèìîäåéñòâèè ïî òèïó II ýíåðãèè ñâÿ- çûâàíèÿ íå òîëüêî ñèëüíî îòëè÷àþòñÿ ìåæäó ñî- áîé, íî è ñàìî ïî ñåáå çíà÷åíèå ýíåðãèè ñâÿ- çûâàíèÿ 2,3 êêàë/ìîëü â ñëó÷àå ÑÀ2 ãîâîðèò î íåâîçìîæíîñòè ñóùåñòâîâàíèÿ ïîäîáíîãî êîìï- ëåêñà. Êîíå÷íî, íåëüçÿ îêîí÷àòåëüíî èñêëþ÷èòü âîçìîæíîñòè ñâÿçûâàíèÿ ïî òèïó II ñîåäèíåíèÿ ÑÀ1, îäíàêî åãî òåðìîäèíàìè÷åñêàÿ âåðîÿòíîñòü íàìíîãî ìåíüøå, ÷åì äëÿ ñâÿçûâàíèÿ ïî òèïó I. Ñóììèðóÿ ðåçóëüòàòû èññëåäîâàíèÿ, ìîæíî óò- âåðæäàòü, ÷òî ïîäîáíî äðóãèì «èíòåðàêòèâíûì» ñàéòàì íà ïîâåðõíîñòè - è -ñóáúåäèíèö òó- áóëèíà (êîëõèöèíîâîìó, âèíáëàñòèíîâîìó/ìýé- òàñèíîâîìó, òàêñàíîâîìó/ýïîòèëîíîâîìó) ñàéò ñâÿçûâàíèÿ äèíèòðîàíèëèíîâûõ ñîåäèíåíèé, îõàðàêòåðèçîâàííûé â íàøèõ ïðåäûäóùèõ ðàáî- òàõ [28–30], ñïîñîáåí ðàñïîçíàâàòü è ñïåöèôè- ÷åñêè ñâÿçûâàòü âåùåñòâà, êàðäèíàëüíî îòëè÷- íûå ïî ñâîåé õèìè÷åñêîé ïðèðîäå è íå èìåþ- ùèå îáùèõ ôàðìàêîôîðíûõ ãðóïï. Îäíàêî ñîå- äèíåíèÿ, ðàçäåëÿþùèå ñàéò âçàèìîäåéñòâèÿ íà ïîâåðõíîñòè áåëêà, äîëæíû ñîäåðæàòü îïðåäå- ëåííûå ñòðóêòóðíûå êîìïîíåíòû, êîòîðûå îá- ëàäàþò ñõîäíîé ýëåêòðîñòàòè÷åñêîé òîïîëîãèåé è, â êîíå÷íîì ñ÷åòå, ÿâëÿþòñÿ îïðåäåëÿþùèìè â ïðîöåññå ìåæìîëåêóëÿðíîãî ðàñïîçíàâàíèÿ (â äàííîì ñëó÷àå òàêèì ñâîéñòâîì îáëàäàþò íèò- ðèëüíàÿ ãðóïïà äèíèòðîàíèëèíîâ è öèàíîãðóï- ïà öèàíîàêðèëàòîâ). Âñëåäñòâèå áîëüøîãî îáúå- ìà äàííûõ, ñâèäåòåëüñòâóþùèõ â ïîëüçó òàêîé ãèïîòåçû, ìîæíî òàêæå ïðåäïîëîæèòü, ÷òî ñâîéñòâîì «ìóëüòèëèãàíäíîñòè» èëè «ìóëüòè- êîìïîíåíòíîñòè» â îòíîøåíèè íèçêîìîëåêó- ëÿðíûõ îðãàíè÷åñêèõ ñîåäèíåíèé ìîãóò îáëà- äàòü è äðóãèå ñàéòû ñïåöèôè÷åñêîãî ñâÿçû- âàíèÿ íèçêîìîëåêóëÿðíûõ ëèãàíäîâ â ìîëå- êóëàõ òóáóëèíîâ. A.Yu. Nyporko, Ya.B. Blume Taras Shevchenko National University of Kyiv, Ukraine Institute of Food Biotechnology and Genomics of National Academy of Sciences of Ukraine, Kyiv E-mail: dfnalex@gmail.com STRUCTURAL MECHANISMS OF INTERACTION OF CYANOACRYLATES WITH PLANT TUBULIN Structural mechanisms underlying the specific binding of cyanoacrylate compounds with tubulin of higher plants have been studied by example of interaction of ethyl- (2Z)-3-amino-2-cyano-4-ethylhex-2-enoate (CA1) and isopropyl-(2Z)-3-amino-2-cyano-4-ethylhex-2-enoate (CA2) with Arabidopsis thaliana -tubulin. It was re- vealed that the cyano group of cyanoacrylates is a func- tional analog of the nitrile group, which determines the processes of specific interaction with plant tubulin for dinitroaniline compounds. Based on the data of spa- tial structure fluctuations, dynamics of hydrogen bonds and interaction energy of the CA1 and CA2, the most probable binding mode for these compounds with plant -tubulin was identified and appropriate site of interac- tion was characterized. Seven out of 10 residues compos- ing this site (Gln-133, Asn-249, Val-250, Asp-251, Val- 252, Asn-253 and Glu-254) are obligatory components of the dinitroanilines’ binding site on the plant -tubulin surface. Thus, the binding site on the -tubulin surface characterized by us is able to recognize and specifically bind the substances which are cardinally different by their chemical nature and have no common pharmaco- phore groups, under the condition of a certain simila- rity of their electrostatic topology. ÑÏÈÑÎÊ ËÈÒÅÐÀÒÓÐÛ 1. Huppatz J.L., Phillips J.N., Rattigan B.M. Cyanoac- rylates. Herbicidal and photosynthetic inhibitory ac- tivity // Agric. Biol. Chem. – 1981. – 45, ¹ 12. – P. 2769–2773. 2. 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