Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues
Phosphorylation of endocytic adaptor ITSN2 that enabled its interaction with the SH2 domains of signaling proteins was recently reported. The aim of this study was to determine whether tissue-specific ITSN2 phosphorylation and subsequent recognition by phosphotyrosine-binding domains could occur in...
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Інститут молекулярної біології і генетики НАН України
2013
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| Назва видання: | Вiopolymers and Cell |
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nasplib_isofts_kiev_ua-123456789-1532132025-02-09T14:30:05Z Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues Диференційне впізнавання білка ITSN2/Ese2 доменами SH2 білків Fyn, Abl1, PLCg1 і PI3KR1 у тканинах миші Дифференциальное узнавание белка ITSN2/Ese2 доменами SH2 белков Fyn, Abl1, PLCg1 и PI3KR1 в тканях мыши Novokhatska, O.V. Pankivskyy, S.V. Dergai, M.V. Tsyba, L.O. Rynditch, A.V. Short Communications Phosphorylation of endocytic adaptor ITSN2 that enabled its interaction with the SH2 domains of signaling proteins was recently reported. The aim of this study was to determine whether tissue-specific ITSN2 phosphorylation and subsequent recognition by phosphotyrosine-binding domains could occur in mouse tissues. Methods. In silico prediction of interaction motifs, expression of recombinant proteins in bacterial system, GST pull-down analysis, immunoblotting. Results. Analysis of phosphoproteomic data demonstrated tyrosine phosphorylation of mouse ITSN2 homologue, Ese2 protein. Scansite service was used to predict binding motifs for the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 within Ese2. Comparison of ITSN2 and Ese2 sequences showed conservation of predicted interaction motifs between human and mouse. GST-fused SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 were obtained and used as phosphorylation «sensors» of tyrosine-based motifs within Ese2 molecule. Binding of Ese2 to the SH2 domains of Fyn and PLCg1 was observed in brain, lung and heart whereas SH2 domains of Abl1 and PI3KR1 interacted with Ese2 in lung and heart only. Conclusions. Differential Ese2/ SH2 interactions in tissues suggest that tissue-specific tyrosine phosphorylation might regulate specific binding of the Ese2 adaptor to the signaling molecules. Нещодавно виявлено фосфорилювання адаптора ендоцитозу ITSN2, що забезпечує впізнавання цього білка SH2-доменами білків, залучених до передачі мітогенного сигналу. Метою цієї роботи було перевірити, чи має взаємодія ITSN2 з SH2-вмісними біл- ками тканиноспецифічний характер. Методи. Передбачення мотивів взаємодії in silico, експресія білків у бактерійній системі та культурі клітин ссавців, преципітація з використанням білків, злитих з GST. Результати. Дані фосфопротеомних досліджень свідчать про фосфорилювання тирозинових залишків гомолога ITSN2 миші, білка Ese2. За допомогою сервісу Scansite у складі Ese2 передбачено мотиви взаємодії з доменами SH2 білків Fyn, Abl1, PLCg1 і PI3KR1. Порівняння послідовностей інтерсектинів людини та миші показало консервативність передбачених мотивів. Отримано злиті з GST домени SH2 білків Fyn, Abl1, PLCg1 і PI3KR1, які використано для преципітації білка Ese2 з лізатів мозку, легень і серця миші. Зв’язування Ese2 з доменами SH2 білків Fyn і PLCg1 спостерігали в усіх досліджуваних тканинах, тоді як домени SH2 білків Abl1 і PI3KR1 упізнавали Ese2 лише в легенях та серці. Висновки. Диференційне впізнавання Ese2/SH2 у тканинах дозволяє припустити, що тканиноспецифічне фосфорилювання регулює специфічне зв’язування адапторного білка Ese2 із сигнальними молекулами. Недавно показано фосфорилирование адаптора эндоцитоза ITSN2, опосредующее узнавание этого белка SH2-доменами белков, вовлеченных в передачу митогенного сигнала. Целью этой работы было проверить, имеет ли взаимодействие ITSN2 с SH2- содержащими белками тканеспецифический характер. Методы. Предсказание мотивов взаимодействия in silico, экспрессия белков в бактериальной системе и культуре клеток млекопитающих, преципитация с использованием белков, слитых с GST. Результаты. Данные фосфопротеомных исследований свидетельствуют о фосфорилировании тирозиновых остатков гомолога ITSN2 мыши, белка Ese2. При помощи сервиса Scansite в составе Ese2 предсказано мотивы взаимодействия с доменами SH2 белков Fyn, Abl1, PLCg1 и PI3KR1. Сравнение последовательностей интерсектинов человека и мыши показало консервативность предсказанных мотивов. Получены слитые с GST домены SH2 белков Fyn, Abl1, PLCg1 и PI3KR1, использованных для преципитации белка Ese2 из лизатов головного мозга, легких и сердца мыши. Связывание Ese2 с доменами SH2 белков Fyn и PLCg1 наблюдали во всех исследованных тканях, тогда как домены SH2 белков Abl1 и PI3KR1 узнавали Ese2 только в тканях легких и сердца. Выводы. Дифференциальное узнавание Ese2/SH2 в тканях позволяет предположить, что тканеспецифическое фосфорилирование опосредует специфическое связывание адаптерного белка Ese2 с сиг- нальными молекулами. 2013 Article Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues / O.V. Novokhatska, S.V. Pankivskyy, M.V. Dergai, L.O. Tsyba, A.V. Rynditch // Вiopolymers and Cell. — 2013. — Т. 29, №. 5. — С. 424-427. — Бібліогр.: 10 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000834 https://nasplib.isofts.kiev.ua/handle/123456789/153213 577.155.2 en Вiopolymers and Cell application/pdf Інститут молекулярної біології і генетики НАН України |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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DSpace DC |
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English |
| topic |
Short Communications Short Communications |
| spellingShingle |
Short Communications Short Communications Novokhatska, O.V. Pankivskyy, S.V. Dergai, M.V. Tsyba, L.O. Rynditch, A.V. Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues Вiopolymers and Cell |
| description |
Phosphorylation of endocytic adaptor ITSN2 that enabled its interaction with the SH2 domains of signaling proteins was recently reported. The aim of this study was to determine whether tissue-specific ITSN2 phosphorylation and subsequent recognition by phosphotyrosine-binding domains could occur in mouse tissues. Methods. In silico prediction of interaction motifs, expression of recombinant proteins in bacterial system, GST pull-down analysis, immunoblotting. Results. Analysis of phosphoproteomic data demonstrated tyrosine phosphorylation of mouse ITSN2 homologue, Ese2 protein. Scansite service was used to predict binding motifs for the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 within Ese2. Comparison of ITSN2 and Ese2 sequences showed conservation of predicted interaction motifs between human and mouse. GST-fused SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 were obtained and used as phosphorylation «sensors» of tyrosine-based motifs within Ese2 molecule. Binding of Ese2 to the SH2 domains of Fyn and PLCg1 was observed in brain, lung and heart whereas SH2 domains of Abl1 and PI3KR1 interacted with Ese2 in lung and heart only. Conclusions. Differential Ese2/ SH2 interactions in tissues suggest that tissue-specific tyrosine phosphorylation might regulate specific binding of the Ese2 adaptor to the signaling molecules. |
| format |
Article |
| author |
Novokhatska, O.V. Pankivskyy, S.V. Dergai, M.V. Tsyba, L.O. Rynditch, A.V. |
| author_facet |
Novokhatska, O.V. Pankivskyy, S.V. Dergai, M.V. Tsyba, L.O. Rynditch, A.V. |
| author_sort |
Novokhatska, O.V. |
| title |
Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues |
| title_short |
Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues |
| title_full |
Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues |
| title_fullStr |
Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues |
| title_full_unstemmed |
Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues |
| title_sort |
differential recognition of itsn2/ese2 by the sh2 domains of fyn, abl1, plcg1 and pi3kr1 in mouse tissues |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2013 |
| topic_facet |
Short Communications |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/153213 |
| citation_txt |
Differential recognition of ITSN2/Ese2 by the SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 in mouse tissues / O.V. Novokhatska, S.V. Pankivskyy, M.V. Dergai, L.O. Tsyba, A.V. Rynditch // Вiopolymers and Cell. — 2013. — Т. 29, №. 5. — С. 424-427. — Бібліогр.: 10 назв. — англ. |
| series |
Вiopolymers and Cell |
| work_keys_str_mv |
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2025-11-26T20:28:06Z |
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2025-11-26T20:28:06Z |
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| fulltext |
SHORT COMMUNICATIONS
UDC 577.155.2
Differential recognition of ITSN2/Ese2 by
the SH2 domains of Fyn, Abl1, PLCg1 and
PI3KR1 in mouse tissues
O. V. Novokhatska, S. V. Pankivskyy, M. V. Dergai, L. O. Tsyba, A. V. Rynditch
State Key Laboratory of Molecular and Cellular Biology,
Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
olga.novokhatska@gmail.com
Phosphorylation of endocytic adaptor ITSN2 that enabled its interaction with the SH2 domains of signaling
proteins was recently reported. The aim of this study was to determine whether tissue-specific ITSN2
phosphorylation and subsequent recognition by phosphotyrosine-binding domains could occur in mouse tissues.
Methods. In silico prediction of interaction motifs, expression of recombinant proteins in bacterial system, GST
pull-down analysis, immunoblotting. Results. Analysis of phosphoproteomic data demonstrated tyrosine phos-
phorylation of mouse ITSN2 homologue, Ese2 protein. Scansite service was used to predict binding motifs for the
SH2 domains of Fyn, Abl1, PLCg1 and PI3KR1 within Ese2. Comparison of ITSN2 and Ese2 sequences showed
conservation of predicted interaction motifs between human and mouse. GST-fused SH2 domains of Fyn, Abl1,
PLCg1 and PI3KR1 were obtained and used as phosphorylation «sensors» of tyrosine-based motifs within Ese2
molecule. Binding of Ese2 to the SH2 domains of Fyn and PLCg1 was observed in brain, lung and heart whereas
SH2 domains of Abl1 and PI3KR1 interacted with Ese2 in lung and heart only. Conclusions. Differential Ese2/
SH2 interactions in tissues suggest that tissue-specific tyrosine phosphorylation might regulate specific binding
of the Ese2 adaptor to the signaling molecules.
Keywords: ITSN2/Ese2, tyrosine phosphorylation, SH2 domain, tissue-specific interactions.
Introduction. Intersectin 1 and 2, ITSN1 and ITSN2,
are endocytic adaptors that mediate assembly of multi-
protein complexes during clathrin-mediated endocyto-
sis. Both proteins have the same domain composition
and are widely expressed in tissues (reviewed in [1, 2]).
Recently we performed functional comparison of ITSNs
and showed that ITSN2 is tyrosine phosphorylated in
contrast to ITSN1. Phosphorylated form of ITSN2 was
recognized by the SH2 domains of signaling proteins in
HeLa cells [3]. According to the phosphoproteomic da-
ta and in silico prediction, nine tyrosine residues of
ITSN2 can be modified and each recognized by several
partners [4, 5]. We suggest that in tissues, modification
of different residues could allow binding of distinct pro-
tein partners. The aim of this study was to verify this hy-
pothesis using mouse model.
Materials and methods. In silico prediction of bin-
ding motifs for SH2 domains was performed using
Scansite service [4].
Expression constructs and antibodies. The GST-
SH2 domains of Fyn, PLCg1, Abl1 and PI3KR1 were
described previously [3]. Polyclonal antibodies against
the CCR of ITSN2 were produced in rabbits immuni-
zed with the recombinant His-tagged protein compri-
sing amino acid residues 349–499 of ITSN2.
Binding assays and Western blot analysis. The re-
combinant GST-fused proteins were expressed in Es-
cherichia coli and affinity purified using Sepharose 4B
(«GE Healthcare», USA) according to the manufactu
rer’s instructions. Lysates of mouse tissues were prepa-
424
ISSN 0233–7657. Biopolymers and Cell. 2013. Vol. 29. N 5. P. 424–427 doi: 10.7124/bc.000834
� Institute of Molecular Biology and Genetics, NAS of Ukraine, 2013
425
DIFFERENTIAL RECOGNITION OF ITSN2/ESE2 BY THE SH2 DOMAINS
red in extraction buffer containing 20 mM Tris-HCl,
pH 7.5, 0.5 % NP40, 150 mM NaCl, 10 % glycerol,
1 mM Na3VO4 and protease inhibitor cocktail («Ro-
che», France). For pull-down assays, immobilized on
beads GST-SH2 domains were incubated with cell lysa-
tes for 1 h at 4 °C. The beads were then washed with ex-
traction buffer three times at 4 °C and boiled in Laem-
mli buffer (150 mM Tris-HCl, pH 6.8, 2.5 % glycerol,
10 % SDS, 3 % �-mercaptoethanol and 0.5 % bromo-
phenol blue).
Proteins were resolved in SDS-PAGE, transferred
to nitrocellulose membranes («Bio-Rad», USA) and
probed with appropriate antibodies for 1 h at room tem-
perature. Detection was performed using ECL reagents.
Chemiluminescence was captured with Molecular Ima-
ger ChemiDocTM XRS+ («Bio-Rad»).
Results and discussion. To verify our hypothesis
about the existence of ITSN2 tissue-specific phospho-
rylation/interaction pattern, mouse tissues were used.
The similarity between human ITSN2 and homologous
mouse Ese2 protein is 92 %. Therefore we used previ-
ously obtained anti-ITSN2 antibodies [3] that were
able to detect major short and long Ese2 isoforms in
mouse tissue lysates. We tested binding of SH2 domains
of Fyn and Abl1 kinases as well as phospholipase C gam-
ma 1 (PLCg1) and regulatory subunit of PI3K (PI3KR1)
to Ese2 from mouse tissues.
Using Scansite service the binding motifs for these
SH2 domains were predicted within Ese2 sequence
(Table). Phosphorylation of Y554 and Y922 of Ese2
protein was demonstrated by phosphoproteomic stu-
dies (Figure, A) [5].
For all proteins except for Fyn kinase conserved bet-
ween human and mouse tyrosine-based interaction
motifs were predicted within Ese2.
Affinity purified GST-SH2 domains of Fyn, PLCg1,
Abl1 and PI3KR1 were used to pull down Ese2 from
mouse brain, lung and heart lysates. In mouse lung and
heart lysates Ese2 was recognized by all tested SH2 do-
mains (Figure, B, left and middle panels). Only the SH2
domains of Fyn and PLCg1 were able to pull down
Ese2 from mouse brain lysate (Figure, B, right panel).
Obtained data indicate that in various tissues phos-
phorylation of distinct tyrosine residues within ITSN2
A
B
554
Ese2
ITSN2
EH1 EH2 SH3A DH PH C2SH3B SH3C SH3D SH3E
922
Short isoform Long isoform C-terminal extension
WB: anti-ITSN2
Coomassie stauning
GST �
GST-Sh2 doamains �
Ese2-L �
Ese2-S �
kDa
� 170
� 130
� 43
� 34
� 26
PI3KR1-NGST
Fyn
PLCg1
AblIInput
PI3KR1-NGST
Fyn
PLCg1
AblIInput
PI3KR1-NGST
Fyn
PLCg1
AblIInput
HeartLung Brain
Ese2 is differentially recognized by various SH2 domains in mouse brain, lung and heart: A – schematic representation of ITSN2/Ese2 domain
organization and distribution of tyrosine residues (shown as black boxes). Tyrosine residues that are phosphorylated according to phosphoproteo-
mic data are indicated by circles. The number above each circle indicates the position of the residue within Ese2 protein; B – Bacterially expressed
and affinity purified GST-SH2 domains of Fyn, Abl1, PI3KR1, PLCg1 or GST alone (control) were bound to glutathione beads and used as bait to
pull down Ese2. For in vitro binding assays GST-fused proteins were incubated with lysates of mouse brain, heart and lung. Bound proteins were
analyzed by Western blotting using antibodies against ITSN2. GST-fused proteins were visualized by Coomassie staining. PI3KR1-N corres-
ponds to the N-terminal SH2 domain of PI3KR1. Ese2-S/L, short/long isoform of Ese2; WB, Western blotting
could occur and demonstrate recognition of ITSN2 by
particular SH2-containing proteins in a tissue-specific
manner. Recent studies focused on investigation of tis-
sue-specific phosphorylation of proteins in mouse tis-
sues indicated that mainly phosphoproteins are widely
expressed [6].
It was suggested that tissue-dependent phospho-
rylation serves for regulation of ubiquitous proteins and
tissue-specific phenotypes are achieved by ubiquitous
pathways. Expression of ITSN2 major isoforms, short
and long, was demonstrated in many human and mouse
tissues [1, 2]. The role of ITSN2 in various cell-specific
processes such as induction of T cell receptor endocyto-
sis, formation of lumen during epithelial morphogenesis,
regulation of actin cytoskeleton dynamics during gast-
rulation and caveolae endocytosis in endothelial cells is
established [7–10].
The discovered tissue-dependent recognition of
ITSN2 by the SH2 domains of signaling proteins could
contribute to ITSN2 function in reported or not yet stu-
died cell type-specific processes.
Acknowledgments. We thank Dr. Larysa Mace-
wicz for providing mouse tissues. This work was par-
tially supported by target complex interdisciplinary pro-
gram of scientific researches of NAS of Ukraine «Fun-
damentals of molecular and cell biotechnologies».
Î. Â. Íîâîõàöüêà, Ñ. Â. Ïàíüê³âñüêèé, Ì. Â. Äåðãàé, Ë. Î. Öèáà,
À. Â. Ðèíäè÷
Äèôåðåíö³éíå âï³çíàâàííÿ á³ëêà ITSN2/Ese2 äîìåíàìè
SH2 á³ëê³â Fyn, Abl1, PLCg1 ³ PI3KR1 ó òêàíèíàõ ìèø³
Ðåçþìå
Íåùîäàâíî âèÿâëåíî ôîñôîðèëþâàííÿ àäàïòîðà åíäîöèòîçó
ITSN2, ùî çàáåçïå÷óº âï³çíàâàííÿ öüîãî á³ëêà SH2-äîìåíàìè
á³ëê³â, çàëó÷åíèõ äî ïåðåäà÷³ ì³òîãåííîãî ñèãíàëó. Ìåòîþ ö³º¿ ðî-
áîòè áóëî ïåðåâ³ðèòè, ÷è ìຠâçàºìîä³ÿ ITSN2 ç SH2-âì³ñíèìè
á³ëêàìè òêàíèíîñïåöèô³÷íèé õàðàêòåð. Ìåòîäè. Ïåðåäáà÷åííÿ
ìîòèâ³â âçàºìî䳿 in silico, åêñïðåñ³ÿ á³ëê³â ó áàêòåð³éí³é ñèñòåì³
òà êóëüòóð³ êë³òèí ññàâö³â, ïðåöèï³òàö³ÿ ç âèêîðèñòàííÿì á³ëê³â,
çëèòèõ ç GST. Ðåçóëüòàòè. Äàí³ ôîñôîïðîòåîìíèõ äîñë³äæåíü
ñâ³ä÷àòü ïðî ôîñôîðèëþâàííÿ òèðîçèíîâèõ çàëèøê³â ãîìîëîãà
ITSN2 ìèø³, á³ëêà Ese2. Çà äîïîìîãîþ ñåðâ³ñó Scansite ó ñêëàä³ Ese2
ïåðåäáà÷åíî ìîòèâè âçàºìî䳿 ç äîìåíàìè SH2 á³ëê³â Fyn, Abl1,
PLCg1 ³ PI3KR1. Ïîð³âíÿííÿ ïîñë³äîâíîñòåé ³íòåðñåêòèí³â ëþ-
äèíè òà ìèø³ ïîêàçàëî êîíñåðâàòèâí³ñòü ïåðåäáà÷åíèõ ìîòèâ³â.
Îòðèìàíî çëèò³ ç GST äîìåíè SH2 á³ëê³â Fyn, Abl1, PLCg1 ³
PI3KR1, ÿê³ âèêîðèñòàíî äëÿ ïðåöèï³òàö³¿ á³ëêà Ese2 ç ë³çàò³â ìîç-
êó, ëåãåíü ³ ñåðöÿ ìèø³. Çâ’ÿçóâàííÿ Ese2 ç äîìåíàìè SH2 á³ëê³â Fyn
³ PLCg1 ñïîñòåð³ãàëè â óñ³õ äîñë³äæóâàíèõ òêàíèíàõ, òîä³ ÿê äî-
ìåíè SH2 á³ëê³â Abl1 ³ PI3KR1 óï³çíàâàëè Ese2 ëèøå â ëåãåíÿõ òà
ñåðö³. Âèñíîâêè. Äèôåðåíö³éíå âï³çíàâàííÿ Ese2/SH2 ó òêàíèíàõ
äîçâîëÿº ïðèïóñòèòè, ùî òêàíèíîñïåöèô³÷íå ôîñôîðèëþâàííÿ
ðåãóëþº ñïåöèô³÷íå çâ’ÿçóâàííÿ àäàïòîðíîãî á³ëêà Ese2 ³ç ñèãíàëü-
íèìè ìîëåêóëàìè.
Êëþ÷îâ³ ñëîâà: ITSN2/Ese2, ôîñôîðèëþâàííÿ òèðîçèíîâèõ çà-
ëèøê³â, äîìåí SH2, òêàíèíîñïåöèô³÷í³ âçàºìî䳿.
Î. Â. Íîâîõàöêàÿ, Ñ. Â. Ïàíüêîâñêèé, Í. Â. Äåðãàé, Ë. À. Öûáà,
À. Â. Ðûíäè÷
Äèôôåðåíöèàëüíîå óçíàâàíèå áåëêà ITSN2/Ese2 äîìåíàìè SH2
áåëêîâ Fyn, Abl1, PLCg1 è PI3KR1 â òêàíÿõ ìûøè
Ðåçþìå
Íåäàâíî ïîêàçàíî ôîñôîðèëèðîâàíèå àäàïòîðà ýíäîöèòîçà
ITSN2, îïîñðåäóþùåå óçíàâàíèå ýòîãî áåëêà SH2-äîìåíàìè áåë-
êîâ, âîâëå÷åííûõ â ïåðåäà÷ó ìèòîãåííîãî ñèãíàëà. Öåëüþ ýòîé
ðàáîòû áûëî ïðîâåðèòü, èìååò ëè âçàèìîäåéñòâèå ITSN2 ñ SH2-
ñîäåðæàùèìè áåëêàìè òêàíåñïåöèôè÷åñêèé õàðàêòåð. Ìåòîäû.
Ïðåäñêàçàíèå ìîòèâîâ âçàèìîäåéñòâèÿ in silico, ýêñïðåññèÿ áåë-
êîâ â áàêòåðèàëüíîé ñèñòåìå è êóëüòóðå êëåòîê ìëåêîïèòàþ-
ùèõ, ïðåöèïèòàöèÿ ñ èñïîëüçîâàíèåì áåëêîâ, ñëèòûõ ñ GST. Ðå-
çóëüòàòû. Äàííûå ôîñôîïðîòåîìíûõ èññëåäîâàíèé ñâèäåòåëü-
ñòâóþò î ôîñôîðèëèðîâàíèè òèðîçèíîâûõ îñòàòêîâ ãîìîëîãà
ITSN2 ìûøè, áåëêà Ese2. Ïðè ïîìîùè ñåðâèñà Scansite â ñîñòàâå
Ese2 ïðåäñêàçàíî ìîòèâû âçàèìîäåéñòâèÿ ñ äîìåíàìè SH2 áåë-
êîâ Fyn, Abl1, PLCg1 è PI3KR1. Ñðàâíåíèå ïîñëåäîâàòåëüíîñòåé
èíòåðñåêòèíîâ ÷åëîâåêà è ìûøè ïîêàçàëî êîíñåðâàòèâíîñòü ïðåä-
ñêàçàííûõ ìîòèâîâ. Ïîëó÷åíû ñëèòûå ñ GST äîìåíû SH2 áåëêîâ
Fyn, Abl1, PLCg1 è PI3KR1, èñïîëüçîâàííûõ äëÿ ïðåöèïèòàöèè
áåëêà Ese2 èç ëèçàòîâ ãîëîâíîãî ìîçãà, ëåãêèõ è ñåðäöà ìûøè.
Ñâÿçûâàíèå Ese2 ñ äîìåíàìè SH2 áåëêîâ Fyn è PLCg1 íàáëþäàëè
âî âñåõ èññëåäîâàííûõ òêàíÿõ, òîãäà êàê äîìåíû SH2 áåëêîâ Abl1
è PI3KR1 óçíàâàëè Ese2 òîëüêî â òêàíÿõ ëåãêèõ è ñåðäöà. Âûâîäû.
Äèôôåðåíöèàëüíîå óçíàâàíèå Ese2/SH2 â òêàíÿõ ïîçâîëÿåò ïðåä-
ïîëîæèòü, ÷òî òêàíåñïåöèôè÷åñêîå ôîñôîðèëèðîâàíèå îïîñðå-
426
NOVOKHATSKA O. V. ET AL.
Fyn PLCg1 Abl1 PI3KR1
– – – Y39
– – – Y194
– Y554 Y554 –
– Y931 – –
Y949 – – –
– Y974 – –
– Y1021 Y1021 Y1124
– – – Y1168
– – – Y1212
– – – Y1534
Y1584 – – –
Ese2 tyrosine residues within predicted SH2-binding motifs
äóåò ñïåöèôè÷åñêîå ñâÿçûâàíèå àäàïòåðíîãî áåëêà Ese2 ñ ñèã-
íàëüíûìè ìîëåêóëàìè.
Êëþ÷åâûå ñëîâà: ITSN2/Ese2, ôîñôîðèëèðîâàíèå òèðîçèíî-
âûõ îñòàòêîâ, äîìåí SH2, òêàíåñïåöèôè÷åñêèå âçàèìîäåéñòâèÿ.
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Received 15.06.13
427
DIFFERENTIAL RECOGNITION OF ITSN2/ESE2 BY THE SH2 DOMAINS
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