TY - JOUR A1 - Mertens, Monique A1 - Hilsch, Malte A1 - Haralampiev, Ivan A1 - Volkmer, Rudolf A1 - Wessig, Pablo A1 - Müller, Peter T1 - Synthesis and characterization of a new Bifunctionalized, Fluorescent, and Amphiphilic molecule for recruiting SH-Containing molecules to membranes JF - ChemBioChem N2 - This study describes the synthesis and characterization of an amphiphilic construct intended to recruit SH-containing molecules to membranes. The construct consists of 1)an aliphatic chain to enable anchoring within membranes, 2)a maleimide moiety to react with the sulfhydryl group of a soluble (bio)molecule, and 3)a fluorescence moiety to allow the construct to be followed by fluorescence spectroscopy and microscopy. It is shown that the construct can be incorporated into preformed membranes, thus allowing application of the approach with biological membranes. The close proximity between the fluorophore and the maleimide moiety within the construct causes fluorescence quenching. This allows monitoring of the reaction with SH-containing molecules by measurement of increases in fluorescence intensity and lifetime. Notably, the construct distributes into laterally ordered membrane domains of lipid vesicles, which is probably triggered by the length of its membrane anchor. The advantages of the new construct can be employed for several biological, biotechnological, and medicinal applications. KW - DBD dyes KW - fatty acids KW - liposomes KW - maleimide KW - membranes KW - palmitoylation Y1 - 2018 U6 - https://doi.org/10.1002/cbic.201800268 SN - 1439-4227 SN - 1439-7633 VL - 19 IS - 15 SP - 1643 EP - 1647 PB - Wiley-VCH CY - Weinheim ER - TY - GEN A1 - Memczak, Henry A1 - Lauster, Daniel A1 - Kar, Parimal A1 - Di Lella, Santiago A1 - Volkmer, Rudolf A1 - Knecht, Volker A1 - Herrmann, Andreas A1 - Ehrentreich-Förster, Eva A1 - Bier, Frank Fabian A1 - Stöcklein, Walter F. M. T1 - Anti-hemagglutinin antibody derived lead peptides for inhibitors of influenza virus binding T2 - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - Antibodies against spike proteins of influenza are used as a tool for characterization of viruses and therapeutic approaches. However, development, production and quality control of antibodies is expensive and time consuming. To circumvent these difficulties, three peptides were derived from complementarity determining regions of an antibody heavy chain against influenza A spike glycoprotein. Their binding properties were studied experimentally, and by molecular dynamics simulations. Two peptide candidates showed binding to influenza A/Aichi/2/68 H3N2. One of them, termed PeB, with the highest affinity prevented binding to and infection of target cells in the micromolar region without any cytotoxic effect. PeB matches best the conserved receptor binding site of hemagglutinin. PeB bound also to other medical relevant influenza strains, such as human-pathogenic A/California/7/2009 H1N1, and avian-pathogenic A/MuteSwan/Rostock/R901/2006 H7N1. Strategies to improve the affinity and to adapt specificity are discussed and exemplified by a double amino acid substituted peptide, obtained by substitutional analysis. The peptides and their derivatives are of great potential for drug development as well as biosensing. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 536 KW - receptor-binding KW - A viruses KW - neutralizing antibody KW - avian influenza KW - origin KW - neuraminidase KW - invection KW - entry KW - sites KW - identification Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-410872 SN - 1866-8372 IS - 536 ER - TY - JOUR A1 - Memczak, Henry A1 - Lauster, Daniel A1 - Kar, Parimal A1 - Di Lella, Santiago A1 - Volkmer, Rudolf A1 - Knecht, Volker A1 - Herrmann, Andreas A1 - Ehrentreich-Foerster, Eva A1 - Bier, Frank Fabian A1 - Stoecklein, Walter F. M. T1 - Anti-Hemagglutinin Antibody Derived Lead Peptides for Inhibitors of Influenza Virus Binding JF - PLoS one N2 - Antibodies against spike proteins of influenza are used as a tool for characterization of viruses and therapeutic approaches. However, development, production and quality control of antibodies is expensive and time consuming. To circumvent these difficulties, three peptides were derived from complementarity determining regions of an antibody heavy chain against influenza A spike glycoprotein. Their binding properties were studied experimentally, and by molecular dynamics simulations. Two peptide candidates showed binding to influenza A/Aichi/2/68 H3N2. One of them, termed PeB, with the highest affinity prevented binding to and infection of target cells in the micromolar region without any cytotoxic effect. PeB matches best the conserved receptor binding site of hemagglutinin. PeB bound also to other medical relevant influenza strains, such as human-pathogenic A/California/7/2009 H1N1, and avian-pathogenic A/MuteSwan/Rostock/R901/2006 H7N1. Strategies to improve the affinity and to adapt specificity are discussed and exemplified by a double amino acid substituted peptide, obtained by substitutional analysis. The peptides and their derivatives are of great potential for drug development as well as biosensing. Y1 - 2016 U6 - https://doi.org/10.1371/journal.pone.0159074 SN - 1932-6203 VL - 11 SP - 82 EP - 90 PB - PLoS CY - San Fransisco ER - TY - CHAP A1 - Haralampiev, Ivan A1 - Mertens, Monique A1 - Schwarzer, Roland A1 - Herrmann, Andreas A1 - Volkmer, Rudolf A1 - Wessig, Pablo A1 - Müller, Peter T1 - A palmitic acid functionalized with a maleimide group is used to recruit SH-containing peptides to lipid and biological membranes T2 - The FEBS journal Y1 - 2015 SN - 1742-464X SN - 1742-4658 VL - 282 SP - 204 EP - 204 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Haralampiev, Ivan A1 - Mertens, Monique A1 - Schwarzer, Roland A1 - Herrmann, Andreas A1 - Volkmer, Rudolf A1 - Wessig, Pablo A1 - Mueller, Peter T1 - Recruitment of SH-Containing peptides to lipid and biological membranes through the use of a palmitic acid functionalized with a Maleimide Group JF - Angewandte Chemie : a journal of the Gesellschaft Deutscher Chemiker ; International edition N2 - This study presents a novel and easily applicable approach to recruit sulfhydryl-containing biomolecules to membranes by using a palmitic acid which is functionalized with a maleimide group. Notably, this strategy can also be employed with preformed (biological) membranes. The applicability of the assay is demonstrated by characterizing the binding of a Rhodamine-labeled peptide to lipid and cellular membranes using methods of fluorescence spectroscopy, lifetime measurement, and microscopy. Our approach offers new possibilities for preparing biologically active liposomes and manipulating living cells. KW - liposomes KW - maleimide KW - membranes KW - palmitic acid KW - palmitoylation KW - peptides Y1 - 2015 U6 - https://doi.org/10.1002/anie.201408089 SN - 1433-7851 SN - 1521-3773 VL - 54 IS - 1 SP - 323 EP - 326 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Daskalow, Katjana A1 - Boisguerin, Prisca A1 - Jandrig, Burkhard A1 - van Landeghem, Frank K. H. A1 - Volkmer, Rudolf A1 - Micheel, Burkhard A1 - Schenk, Jörg A. T1 - Generation of an antibody against the protein phosphatase 1 inhibitor KEPI and characterization of the epitope N2 - A monoclonal antibody against the potential tumor suppressor kinase-enhanced protein phosphatase 1 (PP1) inhibitor KEPI (PPP1R14C) was generated and characterized. Human KEPI was expressed in Escherichia coli and used to immunize Balb/c mice. Using hybridoma technology, one clone, G18AF8, was isolated producing antibodies which bound specifically to the KEPI protein in ELISA, immunoblotting and flow cytometry. The antibody was also successfully applied to stain KEPI protein in paraffin sections of human brain. The epitope was mapped using peptide array technology and confirmed as GARVFFQSPR. This corresponds to the N-terminal region of KEPI. Amino acid substitution analysis revealed that two residues, F and Q, are essential for binding. Affinity of binding was determined by competitive ELISA as 1 mu M. In Western blot assays testing G18AF8 antibody on brain samples of several species, reactivity with hamster, rat and chicken samples was found, suggesting a broad homology of this KEPI epitope in vertebrates. This antibody could be used in expression studies at the protein level e.g. in tumor tissues. Y1 - 2010 UR - http://ar.iiarjournals.org/ SN - 0250-7005 ER -