Single Point Mutation in Bin/Amphiphysin/Rvs (BAR) Sequence of Endophilin Impairs Dimerization, Membrane Shaping, and Src Homology 3 Domain-mediated Partnership

Gortat, Anna; San-Roman, Mabel Jouve; Vannier, Christian; Schmidt, Anne

doi:10.1074/jbc.m111.325837

Cited by 21 publications

(27 citation statements)

References 48 publications

(57 reference statements)

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“…This hypothesis is supported by previous reports of low dimerization affinities for this protein (16,35,45,46).…”

Section: Resultssupporting

confidence: 88%

“…Our finding of subnanomolar affinity rationalizes previous studies that are consistent with the predominance of N-BAR dimers over monomers at sub/low micromolar concentrations: distance measurements via EPR methods in solution at low micromolar concentrations (17), biochemical analysis of extracts from rat brain (71) and from cultured cells exogenously expressing endophilin A2 (45), and fluorescence microscopy analysis of endophilin A2 and A1 fluorescent protein chimeras in cultured cells (35). An analogous notion has been advanced for the BAR protein sorting nexin 9 (72).…”

Section: Discussionsupporting

confidence: 90%

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Kinetics of Endophilin N-BAR Domain Dimerization and Membrane Interactions

Capraro

Shi

Wang

et al. 2013

Journal of Biological Chemistry

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Background: Endocytosis can involve dimerization and membrane association of the protein endophilin. Results: We found subnanomolar affinity for endophilin N-BAR dimerization. Membrane dissociation is substantially slower than association, and membrane-bound protein density-dependent. Conclusion: Endophilin binds membranes as dimers that subsequently oligomerize. Significance: Our findings illuminate the membrane binding mechanism of endophilin, which is important in understanding the regulation of membrane trafficking events.

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“…This hypothesis is supported by previous reports of low dimerization affinities for this protein (16,35,45,46).…”

Section: Resultssupporting

confidence: 88%

Section: Discussionsupporting

confidence: 90%

Kinetics of Endophilin N-BAR Domain Dimerization and Membrane Interactions

Capraro

Shi

Wang

et al. 2013

Journal of Biological Chemistry

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“…Not as much is known about the dimers of endophilin III. The structural information indicates that endophilin III might have more conformational space to bind to other proteins such as dynamin [4][5][6][7] . [32] .…”

Section: Interactions Of Dynamin I With Endophilin I/ii/ Iiimentioning

confidence: 99%

“…Studies have shown that three types of endophilin exist in rats: endophilin I is expressed only in brain, endophilin II in multiple tissues, and endophilin III mainly in brain, testis and thymus [1][2][3] . Endophilins I and II also form dimers through a coiled-coil domain [4][5][6][7] . All three isoforms are concentrated in presynaptic nerve terminals [8][9][10] , and play important roles in synaptic vesicle recycling in neurons [11,12] .…”

Section: Introductionmentioning

confidence: 99%

Endophilin isoforms have distinct characteristics in interactions with N-type Ca2+ channels and dynamin I

Tian¹,

Zhang²,

Fan

et al. 2012

Neurosci. Bull.

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Objective Formation of the endophilin II-Ca 2+ channel complex is Ca 2+ -dependent in clathrin-mediated endocytosis. However, little is known about whether the other two endophilin isoforms have the same features. The present study aimed to investigate the characteristics of the interactions of all three isoforms with Ca 2+ channels and dynamin I. Methods N-type Ca 2+ channel C-terminal fragments (NCFs) synthesized with a 3 H-leucine-labeled kit, were incubated with endophilin-GST fusion proteins, followed by pull-down assay. Results were counted on a scintillation counter. In addition, the different endophilin isoforms were each co-transfected with dynamin I into 293T cells, followed by flow cytometry and co-immunoprecipitation assay. Immunostaining was performed and an image analysis program was used to evaluate the overlap coefficient of cells expressing endophilin and dynamin I. Results All three isoforms interacted with NCF. Endophilins I and II demonstrated clear Ca 2+ -dependent interactions with NCF, whereas endophilin III did not. Co-immunoprecipitation showed that, compared to endophilin I/II, the interaction between endophilin III and dynamin I was significantly increased. Similar results were obtained from flow cytometry. Furthermore, endophilin III had a higher overlap coefficient with dynamin I in co-transfected 293T cells. Conclusion Endophilin isoforms have distinct characteristics in interactions with NCF and dynamin I. Endophilin III binding to NCF is Ca 2+ -independent, implying that it plays a different role in clathrin-mediated endocytosis.

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“…BAR domains are found as dimers, and the importance of their dimerization is illustrated by experiments where dimerizationimpaired endophilin mutants are unable to bind and bend membranes (14,15). Both homodimerization and heterodimerization have been observed.…”

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confidence: 99%

Identification and Characterization of Rvs162/Rvs167-3, a Novel N-BAR Heterodimer in the Human Fungal Pathogen Candida albicans

et al. 2015

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bMembrane reshaping resides at the core of many important cellular processes, and among its mediators are the BAR (Bin, Amphiphysin, Rvs) domain-containing proteins. We have explored the diversity and function of the Rvs BAR proteins in Candida albicans and identified a novel family member, Rvs167-3 (orf19.1861). We show that Rvs167-3 specifically interacts with Rvs162 to form a stable BAR heterodimer able to bind liposomes in vitro. A second, distinct heterodimer is formed by the canonical BAR proteins Rvs161 and Rvs167. Purified Rvs161/Rvs167 complex also binds liposomes, indicating that C. albicans expresses two functional BAR heterodimers. We used live-cell imaging to localize green fluorescent protein (GFP)-tagged Rvs167-3 and Rvs167 and show that both proteins concentrate in small cortical spots. However, while Rvs167 strictly colocalizes with the endocytic marker protein Abp1, we do not observe any colocalization of Rvs167-3 with sites of endocytosis marked by Abp1. Furthermore, the rvs167-3⌬/⌬ mutant is not defective in endocytosis and strains lacking Rvs167-3 or its partner Rvs162 do not display increased sensitivity to high salt concentrations or decreased cell wall integrity, phenotypes which have been observed for rvs167⌬/⌬ and rvs161⌬/⌬ strains and which are linked to endocytosis defects. Taken together, our results indicate different roles for the two BAR heterodimers in C. albicans: the canonical Rvs161/Rvs167 heterodimer functions in endocytosis, whereas the novel Rvs162/Rvs167-3 heterodimer seems not to be involved in this process. Nevertheless, despite their different roles, our phenotypic analysis revealed a genetic interaction between the two BAR heterodimers, suggesting that they may have related but distinct membrane-associated functions.

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Single Point Mutation in Bin/Amphiphysin/Rvs (BAR) Sequence of Endophilin Impairs Dimerization, Membrane Shaping, and Src Homology 3 Domain-mediated Partnership

Cited by 21 publications

References 48 publications

Kinetics of Endophilin N-BAR Domain Dimerization and Membrane Interactions

Kinetics of Endophilin N-BAR Domain Dimerization and Membrane Interactions

Endophilin isoforms have distinct characteristics in interactions with N-type Ca2+ channels and dynamin I

Identification and Characterization of Rvs162/Rvs167-3, a Novel N-BAR Heterodimer in the Human Fungal Pathogen Candida albicans

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