Synthetic Inhibitors of Proline-Rich Ligand-Mediated Protein-Protein Interaction

Oneyama, Chitose; Agatsuma, Tsutomu; Kanda, Yutaka; Nakano, Hirofumi; Sharma, Sreenath V.; Nakano, Satoshi; Narazaki, Fumie; Tatsuta, Kuniaki

doi:10.1016/s1074-5521(03)00101-7

Cited by 43 publications

(31 citation statements)

References 36 publications

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“…Immunoblotting of cell lysates was performed as previously described (Kotani et al, 2007). Immunoprecipitation were performed as previously described (Oneyama et al, 2003). Briefly, cell lysates were quantified and equal amounts of total cell protein were immunoprecipitated overnight with the indicated antibodies (3 g FLAG antibody and 3 g PI3K antibody).…”

Section: Immunoblotting and Immunoprecipitation Assaymentioning

confidence: 99%

The guanine nucleotide exchange factor Arhgef5 plays crucial roles in Src-induced podosome formation

Kuroiwa

Oneyama

Nada

et al. 2011

Journal of Cell Science

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SummaryPodosomes and invadopodia are actin-rich membrane protrusions that play a crucial role in cell adhesion and migration, and extracellular matrix remodeling in normal and cancer cells. The formation of podosomes and invadopodia is promoted by upregulation of some oncogenic molecules and is closely related to the invasive potential of cancer cells. However, the molecular mechanisms underlying the podosome and invadopodium formation still remain unclear. Here, we show that a guanine nucleotide exchange factor (GEF) for Rho family GTPases (Arhgef5) is crucial for Src-induced podosome formation. Using an inducible system for Src activation, we found that Src-induced podosome formation depends upon the Src SH3 domain, and identified Arhgef5 as a Src SH3-binding protein. RNA interference (RNAi)-mediated depletion of Arhgef5 caused robust inhibition of Src-dependent podosome formation. Overexpression of Arhgef5 promoted actin stress fiber remodeling through activating RhoA, and the activation of RhoA or Cdc42 was required for Src-induced podosome formation. Arhgef5 was tyrosine-phosphorylated by Src and bound to Src to positively regulate its activity. Furthermore, the pleckstrin homology (PH) domain of Arhgef5 was required for podosome formation, and Arhgef5 formed a ternary complex with Src and phosphoinositide 3-kinase when Src and/or Arhgef5 were upregulated. These findings provide novel insights into the molecular mechanisms of podosome and invadopodium formation induced by Src upregulation.

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Section: Immunoblotting and Immunoprecipitation Assaymentioning

confidence: 99%

The guanine nucleotide exchange factor Arhgef5 plays crucial roles in Src-induced podosome formation

Kuroiwa

Oneyama

Nada

et al. 2011

Journal of Cell Science

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“…Interaction of these two mutants with Rho* was detected through binding experiment as described in Fig. 2B. protein interactions are important in many signaling pathways (35,36). In the GRK-2 structure, the conserved proline residues form a hydrophobic patch on the surface and their backbone carbonyl oxygen atoms have the potential to form hydrogenbonding interactions (28).…”

Section: Fig 2 Interaction Of Grks-cc With Rhodopsinmentioning

confidence: 99%

Involvement of the C-terminal Proline-rich Motif of G Protein-coupled Receptor Kinases in Recognition of Activated Rhodopsin

Gan

Ma²,

Deng³

et al. 2004

Journal of Biological Chemistry

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G protein-coupled receptor kinases (GRKs) are a family of serine/threonine kinases that phosphorylate many activated G protein-coupled receptors (GPCRs) and play an important role in GPCR desensitization. Our previous work has demonstrated that the C-terminal conserved region (CC) of GRK-2 participates in interaction with rhodopsin and that this interaction is necessary for GRK-2-mediated receptor phosphorylation (Gan, X. Q., Wang, J. Y., Yang, Q. H., Li, Z., Liu, F., Pei, G., and Li, L. (2000) J. Biol. Chem. 275, 8469 -8474). In this report, we further investigated whether the CC of other GRKs had the same functions and defined the specific sequences in CC that are required for the functions. The CC regions of GRK-1, GRK-2, and GRK-5, representatives of the three subfamilies of GRKs, could bind rhodopsin in vitro and inhibit GRK-2-mediated phosphorylation of rhodopsin, but not a peptide GRK substrate. Through a series of mutagenesis analyses, a proline-rich motif in the CC was identified as the key element involved in the interaction between the CC region and rhodopsin. Point mutations of this motif not only disrupted the interaction of GRK-2 with rhodopsin but also abolished the ability of GRK-2 to phosphorylate rhodopsin. The findings that the CC region of GRKs interact only with the light-activated but not the non-activated rhodopsin and that the Nterminal domain of GRK-2 interacts with rhodopsin in a light-independent manner suggest that the CC region is responsible for the recognition of activated GPCRs in the canonical model.

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“…as an inhibitor of capillary tube formation in human umbilical vein endothelial cells (HUVEC) [8,9]. Since a sugar-free derivative of luminacin C did not lose the biological activity [7], we also prepared the similar structure based on migracin A.…”

Section: Discussionmentioning

confidence: 99%

“…Migracin A has a similar structure of luminacin C. In case of luminacin C, a sugar-free dialdehydederivative was synthesized, and it showed the inhibitory activity on HUVEC tube formation [7]. Therefore, we designed the structure of migracinalas the luminacin analog having ethoxy moiety instead of methoxymoiety.…”

Section: Molecular Design Of Migracinalmentioning

confidence: 99%

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Molecular Design of Sugar-Free Migracin Analog Migracinal That Inhibits Ovarian Cancer Cell Migration and Invasion

Ukaji¹,

Koide²,

Lin³

et al. 2017

Kreat. hir. onkol.

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Introduction. Cancer metastasis consists of several steps including detachment from the primary tumor, migration, invasion, transport in the blood or lymphatic vessels, attachment at the secondary site, and growth of secondary tumor. Migration and invasion areinvolved in the mechanism of all types of cancer metastasis. We previously isolated novel cellular migration inhibitor migracin A and B from a culture filtrate of Streptomyces sp. Migracin A was shown to inhibit IGF-1-mediated cellular migration and invasion in ovarian carcinoma cells. However, it is difficult to prepare large amount of migracin A. Migracin A consists of substituted benzene and an alkylated sugar moiety. In the present research, we have designed and synthesized a simplified dialdehydederivative of migracin called migracinal having no sugar moiety.Material and methods. Migracinal was purchased from Techno Chem Co., Ltd., Tokyo, Japan. Migracinal was prepared from 2,4-dihydroxybenzaldehyde (2,4-DHBA). The structure was confirmed by proton and carbon NMR spectra and ESI mass spectroscopy. The antitumor activity of the new derivative was studied by standard tests under conditions in vitro.Results. Migracinal inhibited cellular migration and invasion in ovarian clear cell carcinoma ES-2 cells. It also inhibited IGF-1 expression as migracin A. Moreover, it induced anoikis rather than apoptosis in ES-2 cells.Conclusions. Migracinal is easier to prepare than migracins, and it may be useful for the mechanistic study and suppression of metastasis.Keywords: Migracinal, Migracin, Migration, Invasion, Anoikis, Ovarian clear cell carcinoma For citation: Ukaji T., Koide N., Lin Y., Banno K., Gantsev Sh., Umezawa K. Molecular design of sugar-free migracin analog migracinal that inhibits ovarian cancer cell migration and invasion. Creative surgery and oncology. 2017;7(4): 16-20. DOI:10.24060/207616-20. DOI:10.24060/ -3093-201716-20. DOI:10.24060/ -7-4-16-20. doi: 10.24060/207616-20. DOI:10.24060/ -3093-2017 16-20 Введение. Процесс метастазирования рака состоит из нескольких этапов: отсо-единение клеток от первичной опухоли, миграцию, инвазию, перемещение в кро-ви или лимфатических сосудах, присоединение и рост вторичной опухоли. Меха-низмы миграции и инвазии универсальны для всех видов рака. Ранее из культуры Streptomyces SP мы выделили Migracin А и В -новые ингибиторы клеточной мигра-ции. Было продемонстрировано как Migracin А ингибирует IGF-1-опосредованную миграцию и инвазию клеток рака яичников. Однако большое количество Migracin А, состоящего из замещенного бензола и алкилированного углеводного фрагмента, син-тезировать трудоемко. В настоящем исследовании мы разработали и синтезировали упрощенное производное диальдегида Migracin, не имеющего углеводного компо-нента, названное Migracinal.Материалы и методы. Migracin приобретался у компании «ТехноХим Co., Лтд» (Токио, Япония). Производное Migracinal получали взаимодействием Migracin с 2,4-дигидроксибензалдегидом. Структура была подтверждена спектрами ЯМР и масс-спектроскопией. Противоопухолевая активно...

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Synthetic Inhibitors of Proline-Rich Ligand-Mediated Protein-Protein Interaction

Cited by 43 publications

References 36 publications

The guanine nucleotide exchange factor Arhgef5 plays crucial roles in Src-induced podosome formation

The guanine nucleotide exchange factor Arhgef5 plays crucial roles in Src-induced podosome formation

Involvement of the C-terminal Proline-rich Motif of G Protein-coupled Receptor Kinases in Recognition of Activated Rhodopsin

Molecular Design of Sugar-Free Migracin Analog Migracinal That Inhibits Ovarian Cancer Cell Migration and Invasion

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