The Experts below are selected from a list of 306 Experts worldwide ranked by ideXlab platform

Jamie H D Cate - One of the best experts on this subject based on the ideXlab platform.

  • role for ribosome associated complex and stress seventy subfamily b rac ssb in Integral Membrane protein translation
    Journal of Biological Chemistry, 2017
    Co-Authors: Ligia Acostasampson, Kristina Doring, Yuping Lin, Bernd Bukau, Gunter Kramer, Jamie H D Cate
    Abstract:

    Targeting of most Integral Membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle, which recognizes a hydrophobic signal sequence near the protein N terminus. Proper folding of these proteins is monitored by the unfolded protein response and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transMembrane helix, the signal sequence recognized by the signal recognition particle, is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these Integral Membrane proteins can divert the nascent protein chains to the ribosome-associated complex and stress-seventy subfamily B chaperones. We also show that quality control of Integral Membrane proteins by ribosome-associated complex-stress-seventy subfamily B couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.

  • role for ribosome associated complex and stress seventy subfamily b rac ssb in Integral Membrane protein translation
    bioRxiv, 2017
    Co-Authors: Ligia Acostasampson, Kristina Doring, Yuping Lin, Bernd Bukau, Gunter Kramer, Jamie H D Cate
    Abstract:

    Targeting of most Integral Membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle (SRP), which recognizes a hydrophobic signal sequence near the protein N-terminus. Proper folding of these proteins is monitored by the unfolded protein response, and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transMembrane helix -- the signal sequence recognized by SRP -- is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these Integral Membrane proteins can divert the nascent protein chains to the ribosome-associated complex (RAC) and Stress-Seventy Subfamily B (Ssb) chaperones. We also show that quality control of Integral Membrane proteins by RAC-Ssb couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.

S. Hiller - One of the best experts on this subject based on the ideXlab platform.

  • Sample Preparation and Technical Setup for NMR Spectroscopy with Integral Membrane Proteins.
    Methods in molecular biology (Clifton N.J.), 2020
    Co-Authors: Hundeep Kaur, Anne Grahl, Jean-baptiste Hartmann, S. Hiller
    Abstract:

    NMR spectroscopy is a method of choice to characterize structure, function, and dynamics of Integral Membrane proteins at atomic resolution. Here, we describe protocols for sample preparation and characterization by NMR spectroscopy of two Integral Membrane proteins with different architecture, the α-helical Membrane protein MsbA and the β-barrel Membrane protein BamA. The protocols describe recombinant expression in E. coli, protein refolding, purification, and reconstitution in suitable Membrane mimetics, as well as key setup steps for basic NMR experiments. These include experiments on protein samples in the solid state under magic angle spinning (MAS) conditions and experiments on protein samples in aqueous solution. Since MsbA and BamA are typical examples of their respective architectural classes, the protocols presented here can also serve as a reference for other Integral Membrane proteins.

  • Solution Nuclear Magnetic Resonance Spectroscopy of Integral Membrane Proteins
    Reference Module in Life Sciences, 2017
    Co-Authors: Raphael Böhm, G. Wagner, S. Hiller
    Abstract:

    Solution nuclear magnetic resonance (NMR) spectroscopy is a generally applicable method for studying structure and function of Integral Membrane proteins at atomic resolution. It provides unique features complementary to other high-resolution techniques. This chapter presents an overview of solution NMR techniques for Integral Membrane proteins, describes their potentials and limitations, and describes successful structure determinations as well as selected functional studies. The discussion includes several practical examples from solution NMR studies of the human voltage-dependent anion channel.

  • Perspectives of solution NMR spectroscopy for structural and functional studies of Integral Membrane proteins
    Molecular Physics, 2013
    Co-Authors: Sina Reckel, S. Hiller
    Abstract:

    This article discusses future perspectives of solution NMR spectroscopy to study structures and functions of Integral Membrane proteins at atomic resolution, based on a review of recent progress in this area. Several selected examples of structure determinations, as well as functional studies of Integral Membrane proteins are highlighted. We expect NMR spectroscopy to make future key scientific contributions to understanding Membrane protein function, in particular for large Membrane protein systems with known three-dimensional structure. Such situations can benefit from the fact that functional NMR studies have substantially less limitations by molecular size than a full de novo structure determination. Therefore, the general potential for NMR spectroscopy to solve biologic key questions associated with Integral Membrane proteins is very promising.

  • 5.7 Solution NMR Spectroscopy of Integral Membrane Proteins
    Comprehensive Biophysics, 2012
    Co-Authors: S. Hiller, G. Wagner
    Abstract:

    Solution nuclear magnetic resonance (NMR) spectroscopy is a generally applicable method for studying structure and function of Integral Membrane proteins at atomic resolution. It provides unique features complementary to other high-resolution techniques. This chapter presents an overview of current solution NMR techniques for Integral Membrane proteins, describes their potentials and limitations, and reviews successful structure determinations as well as selected functional studies. The discussion includes several practical examples from solution NMR studies of the human voltage-dependent anion channel.

Martine Cadene - One of the best experts on this subject based on the ideXlab platform.

  • Mass spectrometry of full-length Integral Membrane proteins to define functionally relevant structural features
    Methods, 2008
    Co-Authors: Guillaume Gabant, Martine Cadene
    Abstract:

    The crystallization and structure determination of Integral Membrane proteins remains a difficult task relying on a good understanding of the behavior of the protein for success. To date, Membrane protein structures are still far outnumbered by soluble protein structures. Mass spectrometry is a powerful and versatile tool offering deep insights into the state of the Integral Membrane protein the structuralist intends to crystallize. With appropriate sample preparation methods, it provides information that can sometimes prove critical at various stages of the structure determination process, from protein expression to model building. Moreover, valuable knowledge is gained when the identified structural features underlie important functional aspects. Electrospray and matrix assisted laser desorption ionization (MALDI) methods, however, face a particular challenge when dealing with Integral Membrane proteins. A MALDI method specifically optimized for Membrane protein analysis is presented here, with detailed information on the sample preparation and deposition, as well as guidelines for domain determination by limited proteolysis. MALDI-time of flight mass spectrometry can be used to do a proper inventory of initiation sites, to tailor a protein to a stable, well-folded form, and to evaluate selenomethionine replacement. These approaches are illustrated with a few examples drawn from the structural biology of ion channels.

  • A robust, detergent-friendly method for mass spectrometric analysis of Integral Membrane proteins.
    Analytical chemistry, 2000
    Co-Authors: Martine Cadene, Brian T. Chait
    Abstract:

    Recent breakthroughs in the high-resolution structural elucidation of ion channels and transporters are prompting a growing interest in methods for characterizing Integral Membrane proteins. These methods are proving extremely valuable in facilitating the production of X-ray diffraction-grade crystals. Here we present a robust and straightforward mass spectrometric procedure that utilizes matrix-assisted laser desorption/ionization to analyze Integral Membrane proteins in the presence of detergents. The utility of this method is illustrated with examples of high-quality mass spectral data obtained from Membrane proteins for which atomic resolution structural studies are ongoing.

Bernd Bukau - One of the best experts on this subject based on the ideXlab platform.

  • role for ribosome associated complex and stress seventy subfamily b rac ssb in Integral Membrane protein translation
    Journal of Biological Chemistry, 2017
    Co-Authors: Ligia Acostasampson, Kristina Doring, Yuping Lin, Bernd Bukau, Gunter Kramer, Jamie H D Cate
    Abstract:

    Targeting of most Integral Membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle, which recognizes a hydrophobic signal sequence near the protein N terminus. Proper folding of these proteins is monitored by the unfolded protein response and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transMembrane helix, the signal sequence recognized by the signal recognition particle, is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these Integral Membrane proteins can divert the nascent protein chains to the ribosome-associated complex and stress-seventy subfamily B chaperones. We also show that quality control of Integral Membrane proteins by ribosome-associated complex-stress-seventy subfamily B couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.

  • role for ribosome associated complex and stress seventy subfamily b rac ssb in Integral Membrane protein translation
    bioRxiv, 2017
    Co-Authors: Ligia Acostasampson, Kristina Doring, Yuping Lin, Bernd Bukau, Gunter Kramer, Jamie H D Cate
    Abstract:

    Targeting of most Integral Membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle (SRP), which recognizes a hydrophobic signal sequence near the protein N-terminus. Proper folding of these proteins is monitored by the unfolded protein response, and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transMembrane helix -- the signal sequence recognized by SRP -- is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these Integral Membrane proteins can divert the nascent protein chains to the ribosome-associated complex (RAC) and Stress-Seventy Subfamily B (Ssb) chaperones. We also show that quality control of Integral Membrane proteins by RAC-Ssb couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.

Kristina Doring - One of the best experts on this subject based on the ideXlab platform.

  • role for ribosome associated complex and stress seventy subfamily b rac ssb in Integral Membrane protein translation
    Journal of Biological Chemistry, 2017
    Co-Authors: Ligia Acostasampson, Kristina Doring, Yuping Lin, Bernd Bukau, Gunter Kramer, Jamie H D Cate
    Abstract:

    Targeting of most Integral Membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle, which recognizes a hydrophobic signal sequence near the protein N terminus. Proper folding of these proteins is monitored by the unfolded protein response and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transMembrane helix, the signal sequence recognized by the signal recognition particle, is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these Integral Membrane proteins can divert the nascent protein chains to the ribosome-associated complex and stress-seventy subfamily B chaperones. We also show that quality control of Integral Membrane proteins by ribosome-associated complex-stress-seventy subfamily B couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.

  • role for ribosome associated complex and stress seventy subfamily b rac ssb in Integral Membrane protein translation
    bioRxiv, 2017
    Co-Authors: Ligia Acostasampson, Kristina Doring, Yuping Lin, Bernd Bukau, Gunter Kramer, Jamie H D Cate
    Abstract:

    Targeting of most Integral Membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle (SRP), which recognizes a hydrophobic signal sequence near the protein N-terminus. Proper folding of these proteins is monitored by the unfolded protein response, and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transMembrane helix -- the signal sequence recognized by SRP -- is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these Integral Membrane proteins can divert the nascent protein chains to the ribosome-associated complex (RAC) and Stress-Seventy Subfamily B (Ssb) chaperones. We also show that quality control of Integral Membrane proteins by RAC-Ssb couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.