Research Projects

PREVIOUS AND CURRENT RESEARCH

Cell membranes compartmentalise metabolic processes and serve as selective barriers for permeation. Therefore, nutrient transport through the plasma membrane is essential to maintain homeostasis within the cell. Many proton-coupled secondary active transporters of the major facilitator superfamily (MFS) are involved in the accumulation of nutrients above extracellular levels. Structural and functional analyses of MFS transporters suggest an alternating-access mechanism for the transport of substrates across the membrane. Here the transporter adopts different conformational states, allowing the substrate binding site to face either side of the membrane. Since MFS transporters are found in all branches of life and often with numerous gene copies, we believe that many if not all of these transporters follow a common transport mechanism.

Proton coupled oligopeptide transporters of the PepT family (also known as the POT or SLC15 family) are responsible for the uptake of a range of different di- and tripeptides, derived from the digestion of dietary proteins, and are highly conserved in all kingdoms of life. The best studied members of this family include the two human peptide transporters, PepT1 and PepT2. Both are also of great pharmacological and pharmaceutical interest as they accept a number of drugs and amino acid-conjugated pro-drugs as substrates. A detailed understanding of the structural basis for substrate recognition can therefore help to convert pharmacologically active compounds into substrates for PepT1 and PepT2 and improve their absorption in the small intestine and subsequent distribution in the body. We therefore study members of the proton-dependent oligopeptide transporter (POT) family in vitro and in vivo using a combination of structural biology methods (X-ray, cryo EM, crystallization chaperones) with biochemical approaches (transport assays, fluorescence spectroscopy, crosslinking, transcriptomics).

FUTURE PROJECTS AND GOALS

• Characterisation of pro- and eukaryotic nutrient transporters in various states of the transport cycle using X-ray crystallography and/or Cryo-EM to decipher a common transport mechanism of MFS transporters.
• Structural and dynamic insights into the binding mode of POTs to peptides, drugs, and inhibitors.
• Insights into structure and function of transport regulators of nutrient transporters by structural systems biology approaches

Integral membrane proteins are a challenging class of proteins in terms of their structural and functional characterisation. Over the years we have developed and established new tools and a workflow for protein production and quality control of membrane proteins including functional assays. Furthermore, we are developing new ways to stabilize integral membrane proteins in vitro upon extraction from their natural environment.

Publications

2024

Custódio TF, Guédez G, Löw C (2024) Transient Co-expression of Membrane Protein Complexes in Mammalian Cells.  Methods Mol Biol. 2810:11-28. doi: 10.1007/978-1-0716-3878-1_2

Kohantorabi M, Ugolotti A, Sochor B, Roessler J, Wagstaffe M, Meinhardt A, Beck EE, Dolling DS, Garcia MB, Creutzburg M, Keller TF, Schwartzkopf M, Vayalil SK, Thuenauer R, Guédez G, Löw C, Ebert G, Protzer U, Hammerschmidt W, Zeidler R, Roth SV, Di Valentin C, Stierle A, Noei H (2024) Light-Induced Transformation of Virus-Like Particles on TiO2. ACS Appl Mater Interfaces. doi: 10.1021/acsami.4c07151.

Jungnickel KEJ, Guelle O, Iguchi M, Dong W, Kotov V, Gabriel F, Debacker C, Dairou J, McCort-Tranchepain I, Laqtom NN, Chan SH, Ejima A, Sato K, Massa López D, Saftig P, Mehdipour AR, Abu-Remaileh M, Gasnier B, Löw C, Damme M (2024) MFSD1 with its accessory subunit GLMP functions as a general dipeptide uniporter in lysosomes. Nat Cell Biol. doi: 10.1038/s41556-024-01436-5

Gabriel F, Spriestersbach L, Fuhrmann A, Jungnickel KEJ, Mostafavi S, Pardon E, Steyaert J, Löw C (2024) Structural basis of thiamine transport and drug recognition by SLC19A3. Nat Commun.15(1):8542. doi: 10.1038/s41467-024-52872-8.

2023

Blanchet CE, Round A, Mertens HDT, Ayyer K, Graewert M, Awel S, Franke D, Dörner K, Bajt S, Bean R, Custódio TF, de Wijn R, Juncheng E, Henkel A, Gruzinov A, Jeffries CM, Kim Y, Kirkwood H, Kloos M, Knoška J, Koliyadu J, Letrun R, Löw C, Makroczyova J, Mall A, Meijers R, Pena Murillo GE, Oberthür D, Round E, Seuring C, Sikorski M, Vagovic P, Valerio J, Wollweber T, Zhuang Y, Schulz J, Haas H, Chapman HN, Mancuso AP, Svergun D (2023) Form factor determination of biological molecules with X-ray free electron laser small-angle scattering (XFEL-SAS). Commun Biol. 6(1):1057. doi: 10.1038/s42003-023-05416-7.

Strauss J, Deng L, Gao S, Toseland A, Bachy C, Zhang C, Kirkham A, Hopes A, Utting R, Joest EF, Tagliabue A, Löw C, Worden AZ, Nagel G, Mock T (2023) Plastid-localized xanthorhodopsin increases diatom biomass and ecosystem productivity in iron-limited surface oceans. Nat Microbiol. doi: 10.1038/s41564-023-01498-5.

Custódio TF, Killer M, Yu D, Puente V, Teufel DP, Pautsch A, Schnapp G, Grundl M, Kosinski J, Löw C (2023) Molecular basis of TASL recruitment by the peptide/histidine transporter 1, PHT1.  Nat Commun. 14(1):5696. doi: 10.1038/s41467-023-41420-5.

Zhang C, Parichenko A, Choi W, Shin S, Panes-Ruiz LA, Belyaev D, Custódio TF, Löw C, Lee JS, Ibarlucea B, Cuniberti G (2023) Sybodies as Novel Bioreceptors toward Field-Effect Transistor-Based Detection of SARS-CoV-2 Antigens. ACS Appl Mater Interfaces. doi: 10.1021/acsami.3c06073

Kotov V, Killer M, Jungnickel KEJ, Lei J, Finocchio G, Steinke J, Bartels K, Strauss J, Dupeux F, Humm AS, Cornaciu I, Márquez JA, Pardon E, Steyaert J, Löw C. (2023) Plasticity of the binding pocket in peptide transporters underpins promiscuous substrate recognition. Cell Rep. 42(8):112831. doi: 10.1016/j.celrep.2023.112831.

Hammarén MM, Luukinen H, Sillanpää A, Remans K, Lapouge K, Custódio T, Löw C, Myllymäki H, Montonen T, Seeger M, Robertson J, Nyman TA, Savijoki K, Parikka M (2023) In vitro and ex vivo proteomics of Mycobacterium marinum biofilms and the development of biofilm-binding synthetic nanobodies. mSystems. e0107322. doi: 10.1128/msystems.01073-22

Guédez G, Loers G, Jeffries CM, Kozak S, Meijers R, Svergun DI,  Schachner M, Löw C (2023)  X-ray structure and function of fibronectin domains two and three of the neural cell adhesion molecule L1. The FASEB Journal. 37:e22823. doi:10.1096/fj.202201511R

Kohantorabi M, Wagstaffe M, Creutzburg M, Ugolotti A, Kulkarni S, Jeromin A, Krekeler T, Feuerherd M, Herrmann A, Ebert G, Protzer U, Guédez G, Löw C, Thuenauer R, Schlueter C, Gloskovskii A, Keller TF, Di Valentin C, Stierle A, Noei H (2023) Adsorption and Inactivation of SARS-CoV-2 on the Surface of Anatase TiO2(101). ACS Appl Mater Interfaces. doi: 10.1021/acsami.2c22078.

2022

Alder A, Wilcke L, Pietsch E, von Thien H, Pazicky S, Löw C, Mesen-Ramirez P, Bachmann A, Burda PC, Kunick C, Sondermann H, Wilson D, Gilberger TW (2022) Functional inactivation of Plasmodium falciparum glycogen synthase kinase GSK3 modulates erythrocyte invasion and blocks gametocyte maturation. J Biol Chem.102360. doi: 10.1016/j.jbc.2022.102360

Killer M, Finocchio G, Mertens HDT, Svergun DI, Pardon E, Steyaert J, Löw C (2022) Cryo-EM Structure of an Atypical Proton-Coupled Peptide Transporter: Di- and Tripeptide Permease C. Front Mol Biosci. 9:917725. doi: 10.3389/fmolb.2022.917725

Wichers JS, Mesén-Ramírez P, Fuchs G, Yu-Strzelczyk J, Stäcker J, von Thien H, Alder A, Henshall I, Liffner B, Nagel G, Löw C, Wilson D, Spielmann T, Gao S, Gilberger TW, Bachmann A, Strauss J. (2022) PMRT1, a Plasmodium-Specific Parasite Plasma Membrane Transporter, Is Essential for Asexual and Sexual Blood Stage Development. mBio.e0062322. https://doi.org/10.1128/mbio.00623-22

Pazicky S, Werle AA, Lei J, Löw C, Weininger U (2022) Impact of distant peptide substrate residues on enzymatic activity of SlyD. Cell Mol Life Sci. 79(3):138. doi: 10.1007/s00018-022-04179-4.

Pazicky S, Alder A, Mertens H, Svergun DI, Gilberger T, Löw C (2022) N-terminal phosphorylation regulates the activity of Glycogen Synthase Kinase 3 from Plasmodium falciparum. Biochem J. BCJ20210829. doi: 10.1042/BCJ20210829.

Kovermann M, Weininger U, Löw C (2022) Completing the family of human EH domains: Solution structure of the internal EH domain of γ-synergin. Protein Sci. 31(4):811-821. doi: 10.1002/pro.4269.

2021

Killer M, Wald J, Pieprzyk J, Marlovits TC, Löw C (2021) PepT1 and PepT2 reveal mechanistic insights into substrate and drug transport across epithelial membranes. Sci. Adv. 7, eabk3259 doi: 10.1126/sciadv.abk3259

Lorenzo R, Defelipe LA, Aliperti L, Niebling S, Custódio TF, Löw C, Schwarz JJ, Remans K, Craig PO, Otero LH, Klinke S, García-Alai M, Sánchez IE, Alonso LG (2021) Deamidation drives molecular aging of the SARS-CoV-2 spike protein receptor-binding motif. J Biol Chem.101175. doi: 10.1016/j.jbc.2021.101175.

Bartels K, Lasitza-Male T, Hofmann H, Löw C (2021)Single-Molecule FRET of Membrane Transport Proteins. ChemBioChem.1439-4227. doi:10.1002/cbic.202100261

Dimitrova-Paternoga L, Jagtap PKA, Cyrklaff A, Vaishali, Lapouge K, Sehr P, Perez K, Heber S, Löw C, Hennig J, Ephrussi A (2021) Molecular basis of mRNA transport by a kinesin-1-atypical tropomyosin complex. Genes Dev. doi: 10.1101/gad.348443.121.

Wichers JS, Wunderlich J, Heincke D, Pazicky S, Strauss J, Schmitt M, Kimmel J, Wilcke L, Scharf S, von Thien H, Burda PC, Spielmann T, Löw C, Filarsky M, Bachmann A, Gilberger TW (2021) Identification of novel inner membrane complex and apical annuli proteins of the malaria parasite Plasmodium falciparum. Cell Microbiol. e13341. doi: 10.1111/cmi.13341.

Kotov V, Mlynek G, Vesper O, Pletzer M, Wald J, Teixeira-Duarte CM, Celia H, Garcia-Alai M, Nussberger S, Buchanan SK, Morais-Cabral JH, Loew C, Djinovic-Carugo K, Marlovits TC (2021) In-depth interrogation of protein thermal unfolding data with MoltenProt. Protein Sci. 30(1):201-217. doi: 10.1002/pro.3986. PMID: 33140490 

2020

Custódio TF, Das H, Sheward DJ, Hanke L, Pazicky S, Pieprzyk J, Sorgenfrei M, Schroer M, Gruzinov A, Jeffries C, Graewert M, Svergun D, Dobrev N, Remans K, Seeger MA, McInerney GM, Murrell B, Hällberg MB, Löw C (2020) Selection, biophysical and structural analysis of synthetic nanobodies that effectively neutralize SARS-CoV-2. Nat Commun. 11(1):5588. doi: 10.1038/s41467-020-19204-y.

Pazicky S, Dhamotharan K, Kaszuba K, Mertens HDT, Gilberger T, Svergun D, Kosinski J, Weininger U, Löw C (2020) Structural role of essential light chains in the apicomplexan glideosome. Commun Biol.3(1):568. doi: 10.1038/s42003-020-01283-8. PMID: 33051581 

López DM, Kählau L, Jungnickel KEJ, Löw C, Damme M (2020)Characterization of the complex of the lysosomal membrane transporter MFSD1 and its accessory subunit GLMP. FASEB J. 34(11):14695-14709. doi: 10.1096/fj.202000912RR. PMID: 32959924

Lasitza T, Bartels K, Wiggers F, Rosenblum G, Jungwirth J, Hofmann H, Löw C (2020) Membrane Chemistry Tunes the Structure of a Peptide Transporter. Angew Chem Int Ed Engl doi: 10.1002/anie.202008226 PMID: 32744783 

Geiger M, Brown C, Wichers JS, Strauss J, Lill A, Thuenauer R, Liffner B, Wilcke L, Lemcke S, Heincke D, Pazicky S, Bachmann A, Löw C, Wilson DW, Filarsky M, Burda PC, Zhang K, Junop M, Gilberger TW (2020) Structural Insights Into PfARO and Characterization of its Interaction With PfAIP. J Mol Biol. 432(4):878-896. doi: 10.1016/j.jmb.2019.12.024 PMID: 31877322

2019

Wichers JS, Scholz JAM, Strauss J, Witt S, Lil lA, Ehnold LI, Neupert N, Liffner B, Lühken  R, Petter M, Lorenzen S, Wilson DW, Löw C, Lavazec C, Bruchhaus I, Tannich E, Gilberger  TW, Bachmann A (2019) Dissecting the Gene Expression, Localization, Membrane Topology, and Function of the Plasmodium falciparum STEVOR Protein Family. MBio. pii: e01500-19. doi:  https://doi.org/10.1128/mBio.01500-19,

Kotov V, Vesper O, Alai M G, Löw C, Marlovits TC (2019) Moltenprot: A High-Throughput Analysis Platform to Assess Thermodynamic Stability of Membrane Proteins and Complexes. Biophysical Journal 116 (3), 191a. https://doi.org/10.1016/j.bpj.2018.11.1060

Kotov, V, Bartels, K, Veith, K, Josts, I, Subhramanyam, UKT, Günther, C, Labahn, J, Marlovits, TC, Moraes, I, Tidow, H, Löw C, and Garcia-Alai, MM (2019) High-throughput stability screening  for detergent-solubilized membrane proteins. Sci Rep. 17;10379. doi: https://doi.org/10.1038/s41598-019-46686-8,

Ural-Blimke Y, Flayhan A, Strauss J, Rantos V, Bartels K, Nielsen R, Pardon E, Steyaert J, Kosinski J, Quistgaard EM, Löw C (2019) Structure of Prototypic Peptide Transporter DtpA from E. coli in Complex with Valganciclovir Provides Insights into Drug Binding of Human PepT1. J Am Chem Soc.141; 2404-2412. doi: https://doi.org/10.1021/jacs.8b11343,

2018

Pieprzyk J, Pazicky S, Löw C (2018) Transient Expression of Recombinant Membrane-eGFP Fusion Proteins in HEK293 Cells. Methods Mol Biol. 1850:17-31. doi: https://doi.org/10.1007/978-1-4939-8730-6 2,

Molledo MM*, Quistgaard EM, Löw C (2018) Tripeptide binding in a proton-dependent oligopeptide transporter. FEBS Letters 592: 3239-3247. doi: https://doi.org/10.1002/1873-3468.13246,

Hajizadeh NR, Pieprzyk J, Skopintsev P, Flayhan A, Svergun DI, Löw C (2018) Probing the architecture of a multi-PDZ domain protein: structure of PDZK1 in solution. Structure 26: 1522-1533.e5. doi: https://doi.org/10.1016/j.str.2018.07.016,

Molledo MM , Quistgaard EM, Flayhan A, Pieprzyk J, Löw C (2018) Multispecific substrate recognition in a proton-dependent oligopeptide transporter. Structure 26:467-476.e464. doi: https://doi.org/10.1016/j.str.2018.01.005, PMID: 29429879

Flayhan A, Mertens H, Ural-Blimke H, Molledo MM, Svergun DI, Löw C (2018) Saposin lipid nanoparticles: a highly versatile and modular tool to reconstitute membrane proteins. Structure 26:345-355.e345. doi: https://doi.org/10.1016/j.str.2018.01.007,

2017

Veith K, Molledo MM, Hernandez YA, Josts I, Nitsche J, Löw C, Tidow H (2017) Lipid-like peptides can stabilize integral membrane proteins for biophysical and structural studies. ChemBioChem 18:1735-1742. doi: https://doi.org/10.1002/cbic.201700235

Quistgaard EM, Molledo MM, Löw C (2017) Structure determination of a major facilitator peptide transporter: Inward facing PepTSt from Streptococcus thermophilus crystallized in space group P3121. PLoS One, 12:e0173126. doi: https://doi.org/10.1371/journal.pone.0173126

2016

Quistgaard E M, Löw C, Guettou F, Nordlund P (2016) Understanding transport by the major facilitator superfamily (MFS): structures pave the way. Nature Reviews Molecular Cell Biology 17:123-132. doi: https://doi.org/10.1038/nrm.2015.25, PMID: 26758938

Quistgaard EM, Weininger U, Ural-Blimke Y, Modig K, Nordlund P, Akke M, Löw C (2016) Molecular insights into substrate recognition and catalytic mechanism of the chaperone and FKBP peptidyl-prolyl isomerase SlyD. BMC Biology 14:82. doi: https://doi.org/10.1186/s12915-016-0300-3

Frauenfeld J, Löving R, Armache JP, Sonnen A, Guettou F, Moberg P, Zhu L, Jegerschöld C, Flayhan A, Briggs JA, Garoff H, Löw C, Cheng Y,Nordlund P (2016) A saposin-lipoprotein nanoparticle system for membrane proteins. Nature Methods 13:345-351. doi: https://doi.org/10.1038/nmeth.3801

2014