Bio

Honors & Awards


  • AAAS/Science Program for Excellence in Science, AAAS (2012–2014)

Professional Education


  • Doctor, Ludwig Maximilian Universitat Munchen (2010)

Stanford Advisors


Publications

Journal Articles


  • Initial Steps of Photosystem II de Novo Assembly and Preloading with Manganese Take Place in Biogenesis Centers in Synechocystis PLANT CELL Stengel, A., Guegel, I. L., Hilger, D., Rengstl, B., Jung, H., Nickelsen, J. 2012; 24 (2): 660-675

    Abstract

    In the cyanobacterium Synechocystis sp PCC 6803, early steps in thylakoid membrane (TM) biogenesis are considered to take place in specialized membrane fractions resembling an interface between the plasma membrane (PM) and TM. This region (the PratA-defined membrane) is defined by the presence of the photosystem II (PSII) assembly factor PratA (for processing-associated TPR protein) and the precursor of the D1 protein (pD1). Here, we show that PratA is a Mn(2+) binding protein that contains a high affinity Mn(2+) binding site (K(d) = 73 ?M) and that PratA is required for efficient delivery of Mn(2+) to PSII in vivo, as Mn(2+) transport is retarded in pratA(-). Furthermore, ultrastructural analyses of pratA(-) depict changes in membrane organization in comparison to the wild type, especially a semicircle-shaped structure, which appears to connect PM and TM, is lacking in pratA(-). Immunogold labeling located PratA and pD1 to these distinct regions at the cell periphery. Thus, PratA is necessary for efficient delivery of Mn(2+) to PSII, leading to Mn(2+) preloading of PSII in the periplasm. We propose an extended model for the spatial organization of Mn(2+) transport to PSII, which is suggested to take place concomitantly with early steps of PSII assembly in biogenesis centers at the cell periphery.

    View details for DOI 10.1105/tpc.111.093914

    View details for Web of Science ID 000302131000021

    View details for PubMedID 22319052

  • The Na+/L-proline transporter PutP FRONTIERS IN BIOSCIENCE-LANDMARK Jung, H., Hilger, D., Raba, M. 2012; 17: 745-759

    Abstract

    The Na?/L-proline transporter PutP is a member of the Na?/solute symporter family (TC 2A.21, SLC5), which contains several hundred proteins of pro- and eukaryotic origin. Within the family, the capability of L-proline uptake is restricted to proteins of prokaryotes. PutP contributes to the use of L-proline as a nutrient. In addition, the transporter may supply cells with compatible solute during adaptation to osmotic stress. Based on these and other functions, PutP is of significance for various bacteria-host interactions including the virulence of human pathogens. A homology model of Escherichia coli PutP was generated based on the crystal structure of the Vibrio parahaemolyticus Na+/galactose symporter. According to the model, PutP has a core structure of five plus five transmembrane domains forming an inverted repeat similar as originally revealed by the crystal structure of the Na+/leucine transporter LeuT. The homology model is experimentally verified by Cys cross-linking and site-directed spin labeling in combination with electron paramagnetic resonance spectroscopy. The putative sites of Na? and L-proline binding are described, and a putative transport mechanism is discussed.

    View details for DOI 10.2741/3955

    View details for Web of Science ID 000300049600022

    View details for PubMedID 22201772

  • Homology Model of the Na+/Proline Transporter PutP of Escherichia coli and Its Functional Implications JOURNAL OF MOLECULAR BIOLOGY Olkhova, E., Raba, M., Bracher, S., Hilger, D., Jung, H. 2011; 406 (1): 59-74

    Abstract

    Na(+)/solute symporters are essential membrane integrated proteins that couple the flow of Na(+) ions driven by electrochemical Na(+) gradients to the transport of solutes across biological membranes. Here, we used a combination of molecular modeling techniques and evolutionary conservation analysis to construct and validate a first model of the Na(+)/proline symporter PutP of Escherichia coli based on the crystal structure of the bacterial Na(+)/galactose symporter vSGLT. Ligand docking experiments were employed to gain information about residues involved in proline binding. The proposed model is consistent with the available experimental data and was further validated by amino acid substitutions and kinetic and protein chemical analyses. Combination of the results of molecular modeling and functional studies predicts the location and organization of the Na(+) and proline binding sites. Remarkably, as proposed computationally and discovered here experimentally, residues Y140, W244, and Y248 of transmembrane segments 4 and 7 are found to be particularly important for PutP function and suggested to participate in proline binding and/or gating.

    View details for DOI 10.1016/j.jmb.2010.11.045

    View details for Web of Science ID 000287281800006

    View details for PubMedID 21130773

  • Backbone Structure of Transmembrane Domain IX of the Na+/Proline Transporter PutP of Escherichia coli BIOPHYSICAL JOURNAL Hilger, D., Polyhach, Y., Jung, H., Jeschke, G. 2009; 96 (1): 217-225

    Abstract

    The backbone structure is determined by site-directed spin labeling, double electron electron resonance measurements of distances, and modeling in terms of a helix-loop-helix construct for a transmembrane domain that is supposed to line the translocation pathway in the 54.3 kDa Na(+)/proline symporter PutP of Escherichia coli. The conformational distribution of the spin labels is accounted for by a rotamer library. An ensemble of backbone models with a root mean-square deviation of less than 2 A is obtained. These models exhibit a pronounced kink near residue T341, which is involved in substrate binding. The kink may be associated with a hinge that allows the protein to open and close an inwardly oriented cavity.

    View details for DOI 10.1016/j.bpj.2008.09.030

    View details for Web of Science ID 000266376200023

    View details for PubMedID 19134477

  • Protein chemical and EPR spectroscopic approaches to monitor membrane protein structure and dynamics. Bacterial Signaling (Krämer, R., and Jung, K., eds.) Hilger D, Jung H 2009: 247-263
  • Function of transmembrane domain IX in the Na+/proline transporter PutP JOURNAL OF MOLECULAR BIOLOGY Raba, M., Baumgartner, T., Hilger, D., Klempahn, K., Haertel, T., Jung, K., Jung, H. 2008; 382 (4): 884-893

    Abstract

    Selected residues of transmembrane domain (TM) IX were previously shown to play key roles in ligand binding and transport in members of the Na(+)/solute symporter family. Using the Na(+)/proline transporter PutP as a model, a complete Cys scanning mutagenesis of TM IX (positions 324 to 351) was performed here to further investigate the functional significance of the domain. G328, S332, Q345, and L346 were newly identified as important for Na(+)-coupled proline uptake. Placement of Cys at one of these positions altered K(m(pro)) (S332C and L346C, 3- and 21-fold decreased, respectively; Q345C, 38-fold increased), K(0.5(Na+)) (S332C, 13-fold decreased; Q345C, 19-fold increased), and/or V(max) [G328C, S332C, Q345C, and L346C, 3-, 22-, 2-, and 8-fold decreased compared to PutP(wild type), respectively]. Membrane-permeant N-ethylmaleimide inhibited proline uptake into cells containing PutP with Cys at distinct positions in the middle (T341C) and cytoplasmic half of TM IX (C344, L347C, V348C, and S351C) and had little or no effect on all other single Cys PutP variants. The inhibition pattern was in agreement with the pattern of labeling with fluorescein-5-maleimide. In addition, Cys placed into the cytoplasmic half of TM IX (C344, L347C, V348C, and S351C) was protected from fluorescein-5-maleimide labeling by proline while Na(+) alone had no effect. Membrane-impermeant methanethiosulfonate ethyltrimethylammonium modified Cys in the middle (A337C and T341C) and periplasmic half (L331C) but not in the cytoplasmic half of TM IX in intact cells. Furthermore, Cys at the latter positions was partially protected by Na(+) but not by proline. Based on these results, a model is discussed according to which residues of TM IX participate in the formation of ligand-sensitive, hydrophilic cavities in the protein that may reconstitute part of the Na(+) and/or proline translocation pathway of PutP.

    View details for DOI 10.1016/j.jmb.2008.07.070

    View details for Web of Science ID 000260024500006

    View details for PubMedID 18692508

  • Role of Ser-340 and Thr-341 in transmembrane domain IX of the Na+/Proline transporter PutP of Escherichia coli in ligand binding and transport JOURNAL OF BIOLOGICAL CHEMISTRY Hilger, D., Bohm, M., Hackmann, A., Jung, H. 2008; 283 (8): 4921-4929

    Abstract

    The Na+/solute symporter family comprises more than 400 members of pro- and eukaryotic origin. Using the Na+/proline transporter PutP of Escherichia coli as a model, the role of two conserved residues, Ser-340 and Thr-341, is investigated to obtain insights into the mechanism of transport catalyzed by members of this family. Substitution of these amino acids alters the transport kinetics of cells and proteoliposomes containing the PutP variants significantly. In particular, the apparent affinities for Na+ and Li+ are reduced by 2 orders of magnitude or more. Also proline binding is affected, albeit to a lesser extent than ion binding. Thereby, the presence of a hydroxyl group at position 341 is essential for high affinity ligand binding. Furthermore, Cys placed at position 340 or 341 reacts with sulfhydryl reagents of different polarity, indicating accessibility from the water phase. In addition, Cys cross-linking suggests proximity of the residues to other amino acids previously shown to be crucial for ligand binding. For these reasons it is suggested that Ser-340 and Thr-341 are located in a ligand translocation pathway. Furthermore, it is proposed that the side chain of Thr-341 directly participates in Na+ binding.

    View details for DOI 10.1074/jbc.M706741200

    View details for Web of Science ID 000253426500050

    View details for PubMedID 18156179

  • High-resolution structure of a Na+/H+ antiporter dimer obtained by pulsed election paramagnetic resonance distance measurements BIOPHYSICAL JOURNAL Hilger, D., Polyhach, Y., Padan, E., Jung, H., Jeschke, G. 2007; 93 (10): 3675-3683

    Abstract

    Transient or partial formation of complexes between biomacromolecules is a general mechanism used to control cellular functions. Several of these complexes escape structure determination by crystallographic means. We developed a new approach for determining the structure of protein dimers in the native environment (e.g., in the membrane) with high resolution in cases where the structure of the two monomers is known. The approach is based on measurements of distance distributions between spin labels in the range between 2 and 6 nanometers by a pulsed electron paramagnetic resonance technique and explicit modeling of spin label conformations. By applying this method to the membrane protein homodimer of the Na(+)/H(+) antiporter NhaA of Escherichia coli, the structure of the presumably physiological dimer was determined. It reveals two points of contact between the two monomers, with one of them confirming results of earlier cross-linking experiments.

    View details for DOI 10.1529/biophysj.107.109769

    View details for Web of Science ID 000250577700032

    View details for PubMedID 17704177

  • Secondary transport of amino acids in prokaryotes JOURNAL OF MEMBRANE BIOLOGY Jung, H., Pirch, T., Hilger, D. 2006; 213 (2): 119-133

    Abstract

    Amino acid transport is a ubiquitous phenomenon and serves a variety of functions in prokaryotes, including supply of carbon and nitrogen for catabolic and anabolic processes, pH homeostasis, osmoprotection, virulence, detoxification, signal transduction and generation of electrochemical ion gradients. Many of the participating proteins have eukaryotic relatives and are successfully used as model systems for exploration of transporter structure and function. Distribution, physiological roles, functional properties, and structure-function relationships of prokaryotic alpha-amino acid transporters are discussed.

    View details for DOI 10.1007/s00232-006-0880-x

    View details for Web of Science ID 000245834300007

    View details for PubMedID 17417701

  • DeerAnalysis2006 - a Comprehensive Software Package for Analyzing Pulsed ELDOR Data Applied Magnetic Resonance Jeschke G, Chechik V, Ionita P, Godt A, Zimmermann H, Banham J, Timmel CR, Hilger D, Jung H 2006; 30: 473-498
  • Assessing oligomerization of membrane proteins by four-pulse DEER: pH-dependent dimerization of NhaA Na+/H+ antiporter of E-coli BIOPHYSICAL JOURNAL Hilger, D., Jung, H., Padan, E., Wegener, C., Vogel, K. P., Steinhoff, H. J., Jeschke, G. 2005; 89 (2): 1328-1338

    Abstract

    The pH dependence of the structure of the main Na(+)/H(+) antiporter NhaA of Escherichia coli is studied by continuous-wave (CW) and pulse electron paramagnetic resonance (EPR) techniques on singly spin-labeled mutants. Residues 225 and 254 were selected for site-directed spin labeling, as previous work suggested that they are situated in domains undergoing pH-dependent structural changes. A well-defined distance of 4.4 nm between residues H225R1 in neighboring molecules is detected by a modulation in double electron-electron resonance data. This indicates that NhaA exists as a dimer, as previously suggested by a low-resolution electron density map and cross-linking experiments. The modulation depth decreases reversibly when pH is decreased from 8 to 5.8. A quantitative analysis suggests a dimerization equilibrium, which depends moderately on pH. Furthermore, the mobility and polarity of the environment of a spin label attached to residue 225 change only slightly with changing pH, while no other changes are detected by CW EPR. As antiporter activity of NhaA changes drastically in the studied pH range, residues 225 and 254 are probably located not in the sensor or ion translocation sites themselves but in domains that convey the signal from the pH sensor to the translocation site.

    View details for DOI 10.1529/biophysj.105.062232

    View details for Web of Science ID 000230822200054

    View details for PubMedID 15894644

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