Volta Phase Plate Cryo-EM Structure of the Human Heterodimeric Amino Acid Transporter 4F2hc-LAT2

International Journal of Molecular Sciences
Jean-Marc Jeckelmann, Dimitrios Fotiadis

Abstract

Heteromeric amino acid transporters (HATs) are protein complexes that catalyze the transport of amino acids across plasma membranes. HATs are composed of two subunits, a heavy and a light subunit, which belong to the solute carrier (SLC) families SLC3 and SLC7. The two subunits are linked by a conserved disulfide bridge. Several human diseases are associated with loss of function or overexpression of specific HATs making them drug targets. The human HAT 4F2hc-LAT2 (SLC3A2-SLC7A8) is specific for the transport of large neutral L-amino acids and specific amino acid-related compounds. Human 4F2hc-LAT2 can be functionally overexpressed in the methylotrophic yeast Pichia pastoris and pure recombinant protein purified. Here we present the first cryo-electron microscopy (cryo-EM) 3D-map of a HAT, i.e., of the human 4F2hc-LAT2 complex. The structure could be determined at ~13 Å resolution using direct electron detector and Volta phase plate technologies. The 3D-map displays two prominent densities of different sizes. The available X-ray structure of the 4F2hc ectodomain fitted nicely into the smaller density revealing the relative position of 4F2hc with respect to LAT2 and the membrane plane.

References

Nov 13, 2001·American Journal of Physiology. Renal Physiology·J ChillarónM Palacín
Feb 11, 2004·Pflügers Archiv : European journal of physiology·François VerreyYoshikatsu Kanai
Apr 23, 2004·Biological Procedures Online·Melanie OhiThomas Walz
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Dec 13, 2006·The Journal of Biological Chemistry·Dietmar WeitzHannelore Daniel
Sep 29, 2009·Journal of Molecular Biology·Fabio CasagrandeDimitrios Fotiadis
Apr 13, 2010·Acta Crystallographica. Section D, Biological Crystallography·P EmsleyK Cowtan
Sep 6, 2011·Nucleic Acids Research·Mikhail A LomizeAndrei L Lomize
Jul 31, 2012·Nature Methods·Sjors H W Scheres, Shaoxia Chen
Sep 25, 2012·Journal of Structural Biology·Sjors H W Scheres
Oct 23, 2012·Protein Expression and Purification·Meritxell CostaManuel Palacín
Mar 20, 2013·Molecular Aspects of Medicine·Dimitrios FotiadisManuel Palacín
Feb 12, 2014·Proceedings of the National Academy of Sciences of the United States of America·Albert RosellDimitrios Fotiadis
Oct 22, 2014·Proceedings of the National Academy of Sciences of the United States of America·Radostin DanevWolfgang Baumeister
Apr 25, 2015·Cell·Yifan ChengThomas Walz
Aug 19, 2015·Journal of Structural Biology·Alexis Rohou, Nikolaus Grigorieff
Mar 8, 2016·ELife·Radostin Danev, Wolfgang Baumeister
Mar 10, 2016·Biophysical Journal·Edward H Egelman
Jan 22, 2017·ELife·Radostin DanevWolfgang Baumeister
Mar 28, 2017·Journal of Structural Biology·Nikhil BiyaniHenning Stahlberg
Apr 4, 2018·Journal of the American Society of Nephrology : JASN·Clara VilchesManuel Palacín

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Citations

Jul 10, 2019·Acta Crystallographica. Section D, Structural Biology·George N ChiduzaS Samar Hasnain
Feb 16, 2020·International Journal of Molecular Sciences·Mariafrancesca Scalise, Cesare Indiveri
Jun 5, 2019·Nature Structural & Molecular Biology·Yongchan LeeOsamu Nureki
Oct 1, 2020·International Journal of Molecular Sciences·Jean-Marc Jeckelmann, Dimitrios Fotiadis
Jun 5, 2020·Science Advances·Renhong YanQiang Zhou
May 12, 2019·Science China. Life Sciences·Dimitrios Fotiadis, Jean-Marc Jeckelmann
Oct 18, 2020·International Journal of Molecular Sciences·Satish KantipudiDimitrios Fotiadis
Oct 6, 2020·Protein Science : a Publication of the Protein Society·Kazumasa OdaOsamu Nureki

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Methods Mentioned

BETA
electron microscopy
X-ray
electrophoresis

Software Mentioned

MotionCor2
ctffind
EPU
RELION
UCSF Chimera
COOT
Chimera
FOCUS

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