Stephan, Milena

[["dc.bibliographiccitation.firstpage","E2042"],["dc.bibliographiccitation.issue","30"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences of the United States of America"],["dc.bibliographiccitation.lastpage","E2049"],["dc.bibliographiccitation.volume","109"],["dc.contributor.author","Krick, Roswitha"],["dc.contributor.author","Busse, Ricarda A."],["dc.contributor.author","Scacioc, Andreea"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Janshoff, Andreas"],["dc.contributor.author","Thumm, Michael"],["dc.contributor.author","Kuehnel, Karin"],["dc.date.accessioned","2018-11-07T09:08:09Z"],["dc.date.available","2018-11-07T09:08:09Z"],["dc.date.issued","2012"],["dc.description.abstract","beta-propellers that bind polyphosphoinositides (PROPPINs), a eukaryotic WD-40 motif-containing protein family, bind via their predicted beta-propeller fold the polyphosphoinositides PtdIns3P and PtdIns(3,5) P-2 using a conserved FRRG motif. PROPPINs play a key role in macroautophagy in addition to other functions. We present the 3.0-angstrom crystal structure of Kluyveromyces lactis Hsv2, which shares significant sequence homologies with its three Saccharomyces cerevisiae homologs Atg18, Atg21, and Hsv2. It adopts a seven-bladed beta-propeller fold with a rare nonvelcro propeller closure. Remarkably, in the crystal structure, the two arginines of the FRRG motif are part of two distinct basic pockets formed by a set of highly conserved residues. In comprehensive in vivo and in vitro studies of ScAtg18 and ScHsv2, we define within the two pockets a set of conserved residues essential for normal membrane association, phosphoinositide binding, and biological activities. Our experiments show that PROPPINs contain two individual phosphoinositide binding sites. Based on docking studies, we propose a model for phosphoinositide binding of PROPPINs."],["dc.description.sponsorship","Deutsche Forschungsgemeinschaft [SFB860]"],["dc.identifier.doi","10.1073/pnas.1205128109"],["dc.identifier.isi","000306992700006"],["dc.identifier.pmid","22753491"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/25961"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Natl Acad Sciences"],["dc.relation.issn","0027-8424"],["dc.title","Structural and functional characterization of the two phosphoinositide binding sites of PROPPINs, a beta-propeller protein family"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1987"],["dc.bibliographiccitation.issue","8"],["dc.bibliographiccitation.journal","Analyst"],["dc.bibliographiccitation.lastpage","1992"],["dc.bibliographiccitation.volume","139"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Kramer, Corinna"],["dc.contributor.author","Steinem, Claudia"],["dc.contributor.author","Janshoff, Andreas"],["dc.date.accessioned","2017-09-07T11:46:55Z"],["dc.date.available","2017-09-07T11:46:55Z"],["dc.date.issued","2014"],["dc.description.abstract","Small molecule sensing is of great importance in pharmaceutical research. While there exist well established screening methods such as EMSA (electrophoretic motility shift assay) or biointeraction chromatography to report on successful binding interactions, there are only a few techniques that allow studying and quantifying the interaction of low molecular weight analytes with a binding partner directly. We report on a binding assay for small molecules based on the reflectivity change of a porous transparent film upon immobilisation of an absorbing substance on the pore walls. The porous matrix acts as a thin optical transparent film to produce interference fringes and accumulates molecules at the inner wall to amplify the sensor response. The benefits and limits of the assay are demonstrated by investigating the binding of biotin labelled with an atto dye to avidin physisorbed within an anodic aluminium oxide membrane."],["dc.identifier.doi","10.1039/c4an00009a"],["dc.identifier.fs","604886"],["dc.identifier.gro","3142213"],["dc.identifier.isi","000333081800024"],["dc.identifier.pmid","24599267"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/11469"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5788"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: DFG [SFB 937 (A08)]"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1364-5528"],["dc.relation.issn","0003-2654"],["dc.rights.access","openAccess"],["dc.title","Binding assay for low molecular weight analytes based on reflectometry of absorbing molecules in porous substrates"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","770"],["dc.bibliographiccitation.issue","5"],["dc.bibliographiccitation.journal","Autophagy"],["dc.bibliographiccitation.lastpage","777"],["dc.bibliographiccitation.volume","9"],["dc.contributor.author","Busse, Ricarda A."],["dc.contributor.author","Scacioc, Andreea"],["dc.contributor.author","Hernandez, Javier M."],["dc.contributor.author","Krick, Roswitha"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Janshoff, Andreas"],["dc.contributor.author","Thumm, Michael"],["dc.contributor.author","Kuehnel, Karin"],["dc.date.accessioned","2018-11-07T09:24:51Z"],["dc.date.available","2018-11-07T09:24:51Z"],["dc.date.issued","2013"],["dc.description.abstract","We characterized phosphoinositide binding of the S. cerevisiae PROPPIN Hsv2 qualitatively with density flotation assays and quantitatively through isothermal titration calorimetry (ITC) measurements using liposomes. We discuss the design of these experiments and show with liposome flotation assays that Hsv2 binds with high specificity to both PtdIns3P and PtdIns(3,5)P-2. We propose liposome flotation assays as a more accurate alternative to the commonly used PIP strips for the characterization of phosphoinositide-binding specificities of proteins. We further quantitatively characterized PtdIns3P binding of Hsv2 with ITC measurements and determined a dissociation constant of 0.67 mu M and a stoichiometry of 2:1 for PtdIns3P binding to Hsv2. PtdIns3P is crucial for the biogenesis of autophagosomes and their precursors. Besides the PROPPINs there are other PtdIns3P binding proteins with a link to autophagy, which includes the FYVE-domain containing proteins ZFYVE1/DFCP1 and WDFY3/ALFY and the PX-domain containing proteins Atg20 and Snx4/Atg24. The methods described could be useful tools for the characterization of these and other phosphoinositide-binding proteins."],["dc.description.sponsorship","[SFB860]"],["dc.identifier.doi","10.4161/auto.23978"],["dc.identifier.isi","000323174100012"],["dc.identifier.pmid","23445924"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/29929"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Landes Bioscience"],["dc.relation.issn","1554-8627"],["dc.title","Qualitative and quantitative characterization of protein-phosphoinositide interactions with liposome-based methods"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","106"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Autophagy"],["dc.bibliographiccitation.lastpage","107"],["dc.bibliographiccitation.volume","9"],["dc.contributor.author","Thumm, Michael"],["dc.contributor.author","Busse, Ricarda A."],["dc.contributor.author","Scacioc, Andreea"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Janshoff, Andreas"],["dc.contributor.author","Kuehnel, Karin"],["dc.contributor.author","Krick, Roswitha"],["dc.date.accessioned","2018-11-07T09:30:44Z"],["dc.date.available","2018-11-07T09:30:44Z"],["dc.date.issued","2013"],["dc.description.abstract","PROPPINs are a family of PtdIns3P and PtdIns(3,5) P-2-binding proteins. The crystal structure now unravels the presence of two distinct phosphoinositide-binding sites at the circumference of the seven bladed beta-propeller. Mutagenesis analysis of the binding sites shows that both are required for normal membrane association and autophagic activities. We identified a set of evolutionarily conserved basic and polar residues within both binding pockets, which are crucial for phosphoinositide binding. We expect that membrane association of PROPPINs is further stabilized by membrane insertions and interactions with other proteins."],["dc.identifier.doi","10.4161/auto.22400"],["dc.identifier.isi","000313077900012"],["dc.identifier.pmid","23069643"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/31376"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Landes Bioscience"],["dc.relation.issn","1554-8627"],["dc.title","It takes two to tango PROPPINs use two phosphoinositide-binding sites"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dc.type.subtype","letter_note"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","e85033"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","PLoS ONE"],["dc.bibliographiccitation.volume","9"],["dc.contributor.author","Vilardi, Fabio"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Clancy, Anne"],["dc.contributor.author","Janshoff, Andreas"],["dc.contributor.author","Schwappach, Blanche"],["dc.date.accessioned","2017-09-07T11:46:54Z"],["dc.date.available","2017-09-07T11:46:54Z"],["dc.date.issued","2014"],["dc.description.abstract","Tail-Anchored (TA) proteins are inserted into the endoplasmic reticulum (ER) membrane of yeast cells via the posttranslational Guided Entry of Tail-Anchored protein (GET) pathway. The key component of this targeting machinery is the ATPase Get3 that docks to the ER membrane by interacting with a receptor complex formed by the proteins Get1 and Get2. A conserved pathway is present in higher eukaryotes and is mediated by TRC40, homolog of Get3, and the recently identified membrane receptors WRB and CAML. Here, we used yeast lacking the GET1 and GET2 genes and substituted them with WRB and CAML. This rescued the growth phenotypes of the GET receptor mutant. We demonstrate that WRB and CAML efficiently recruit Get3 to the ER membrane and promote the targeting of the TA proteins in vivo. Our results show that the membrane spanning segments of CAML are essential to create a functional receptor with WRB and to ensure TA protein membrane insertion. Finally, we determined the binding parameters of TRC40 to the WRB/CAML receptor. We conclude that together, WRB and CAML are not only necessary but also sufficient to create a functional membrane receptor complex for TRC40. The yeast complementation assay can be used to further dissect the structure-function relationship of the WRB/CAML heteromultimer in the absence of endogenous receptor proteins."],["dc.description.sponsorship","Open-Access-Publikatinsfonds 2014"],["dc.identifier.doi","10.1371/journal.pone.0085033"],["dc.identifier.gro","3142201"],["dc.identifier.isi","000329460100086"],["dc.identifier.pmid","24392163"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/9662"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5654"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: University Medicine Gottingen"],["dc.notes.intern","Merged from goescholar"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1932-6203"],["dc.rights","CC BY 4.0"],["dc.rights.uri","https://creativecommons.org/licenses/by/4.0"],["dc.title","WRB and CAML Are Necessary and Sufficient to Mediate Tail-Anchored Protein Targeting to the ER Membrane"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.volume","102"],["dc.contributor.author","Lorenz, Baerbel"],["dc.contributor.author","Oelkers, Marieelen"],["dc.contributor.author","Brand, Christian"],["dc.contributor.author","Kriemen, Ella"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Sunnick, Eva"],["dc.contributor.author","Yueksel, Deniz"],["dc.contributor.author","Kumar, Krishna"],["dc.contributor.author","Werz, Daniel B."],["dc.contributor.author","Janshoff, Andreas"],["dc.date.accessioned","2018-11-07T09:14:14Z"],["dc.date.available","2018-11-07T09:14:14Z"],["dc.date.issued","2012"],["dc.format.extent","427A"],["dc.identifier.isi","000321561203035"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/27360"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Cell Press"],["dc.publisher.place","Cambridge"],["dc.relation.eventlocation","San Diego, CA"],["dc.relation.issn","0006-3495"],["dc.title","Weak Carbohydrate-Carbohydrate Interactions Measured by Colloidal Probe Microscopy"],["dc.type","conference_abstract"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","3326"],["dc.bibliographiccitation.issue","7"],["dc.bibliographiccitation.journal","Journal of the American Chemical Society"],["dc.bibliographiccitation.lastpage","3329"],["dc.bibliographiccitation.volume","134"],["dc.contributor.author","Lorenz, Baerbel"],["dc.contributor.author","Alvarez de Cienfuegos, Luis"],["dc.contributor.author","Oelkers, Marieelen"],["dc.contributor.author","Kriemen, Ella"],["dc.contributor.author","Brand, Christian"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Sunnick, Eva"],["dc.contributor.author","Yueksel, Deniz"],["dc.contributor.author","Kalsani, Venkateshwarlu"],["dc.contributor.author","Kumar, Krishna"],["dc.contributor.author","Werz, Daniel B."],["dc.contributor.author","Janshoff, Andreas"],["dc.date.accessioned","2018-11-07T09:13:20Z"],["dc.date.available","2018-11-07T09:13:20Z"],["dc.date.issued","2012"],["dc.description.abstract","The multivalent carbohydrate-carbohydrate interaction between membrane-anchored epitopes derived from the marine sponge Microciona prolifera has been explored by colloidal probe microscopy. An in situ coupling of sulfated and non-sulfated disaccharides to membrane-coated surfaces was employed to mimic native cell cell contacts. The dynamic strength of the homomeric self-association was measured as a function of calcium ions and loading rate. A deterministic model was used to estimate the basic energy landscape and number of participating bonds in the contact zone."],["dc.identifier.doi","10.1021/ja210304j"],["dc.identifier.isi","000301084700004"],["dc.identifier.pmid","22296574"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/27150"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Amer Chemical Soc"],["dc.relation.issn","0002-7863"],["dc.title","Model System for Cell Adhesion Mediated by Weak Carbohydrate-Carbohydrate Interactions"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1366"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Analytical Chemistry"],["dc.bibliographiccitation.lastpage","1371"],["dc.bibliographiccitation.volume","86"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Mey, Ingo"],["dc.contributor.author","Steinem, Claudia"],["dc.contributor.author","Janshoff, Andreas"],["dc.date.accessioned","2017-09-07T11:46:32Z"],["dc.date.available","2017-09-07T11:46:32Z"],["dc.date.issued","2014"],["dc.description.abstract","The passage of solutes across a lipid membrane plays a central role in many cellular processes. However, the investigation of transport processes remains a serious challenge in pharmaceutical research, particularly the transport of uncharged cargo. While translocation reactions of ions across cell membranes is commonly measured with the patch-clamp, an equally powerful screening method for the transport of uncharged compounds is still lacking. A combined setup for reflectometric interference spectroscopy (RIfS) and fluorescence microscopy measurements is presented that allows one to investigate the passive exchange of uncharged compounds across a free-standing membrane. Pore-spanning lipid membranes were prepared by spreading giant 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicles on porous anodic aluminum oxide (AAO) membranes, creating sealed attoliter-sized compartments. The time-resolved leakage of different dye molecules (pyranine and crystal violet) as well as avidin through melittin induced membrane pores and defects was investigated."],["dc.identifier.doi","10.1021/ac4020726"],["dc.identifier.gro","3142186"],["dc.identifier.isi","000331014800011"],["dc.identifier.pmid","24377291"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5488"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: [SFB 937 (A8)]"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1520-6882"],["dc.relation.issn","0003-2700"],["dc.title","Combining Reflectometry and Fluorescence Microscopy: An Assay for the Investigation of Leakage Processes across Lipid Membranes"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","7031"],["dc.bibliographiccitation.issue","20"],["dc.bibliographiccitation.journal","Journal of the American Chemical Society"],["dc.bibliographiccitation.lastpage","7039"],["dc.bibliographiccitation.volume","131"],["dc.contributor.author","Mey, Ingo"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Schmitt, Eva K."],["dc.contributor.author","Müller, Martin Michael"],["dc.contributor.author","Ben Amar, Martine"],["dc.contributor.author","Steinem, Claudia"],["dc.contributor.author","Janshoff, Andreas"],["dc.date.accessioned","2017-09-07T11:47:28Z"],["dc.date.available","2017-09-07T11:47:28Z"],["dc.date.issued","2009"],["dc.description.abstract","The mechanical behavior of lipid bilayers; spanning the pores of highly ordered porous silicon substrates was scrutinized by local indentation experiments as a function of surface functionalization, lipid composition, solvent content, indentation velocity, and pore radius. Solvent-containing nano black lipid membranes (nano-BLMs) as well as solvent-free pore-spanning bilayers were imaged by fluorescence and atomic force microscopy prior to force curve acquisition, which allows distinguishing between membrane-covered and uncovered pores. Force indentation curves on pore-spanning bilayers attached to functionalized hydrophobic porous silicon substrates reveal a predominately linear response that is mainly attributed to prestress in the membranes. This is in agreement with the observation that indentation leads to membrane lysis well below 5% area dilatation. However, membrane bending and lateral tension dominate over prestress and stretching if solvent-free supported membranes obtained from spreading giant liposomes on hydrophilic porous silicon are indented. An elastic regime diagram is presented that readily allows determining the dominant contribution to the mechanical response upon indentation as a function of load and pore radius."],["dc.identifier.doi","10.1021/ja809165h"],["dc.identifier.gro","3143115"],["dc.identifier.isi","000266484700046"],["dc.identifier.pmid","19453196"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/594"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: DFG [JA 963/8-1, STE 884/9-1]"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0002-7863"],["dc.title","Local Membrane Mechanics of Pore-Spanning Bilayers"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.journal","European Biophysics Journal"],["dc.bibliographiccitation.volume","40"],["dc.contributor.author","Stephan, Milena"],["dc.contributor.author","Janshoff, Andreas"],["dc.date.accessioned","2018-11-07T08:53:34Z"],["dc.date.available","2018-11-07T08:53:34Z"],["dc.date.issued","2011"],["dc.format.extent","60"],["dc.identifier.isi","000293637300083"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/22444"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Springer"],["dc.publisher.place","New york"],["dc.relation.eventlocation","Budapest, HUNGARY"],["dc.relation.issn","0175-7571"],["dc.title","Development of a biomembrane sensor based on reflectometry"],["dc.type","conference_abstract"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]