The Different Effects of Substrates and Nucleotides on the Complex Formation of ABC Transporters

Structure
Francesco FiorentinoCarol V Robinson

Abstract

The molybdate importer (ModBC-A of Archaeoglobus fulgidus) and the vitamin B12 importer (BtuCD-F of Escherichia coli) are members of the type I and type II ABC importer families. Here we study the influence of substrate and nucleotide binding on complex formation and stability. Using native mass spectrometry we show that the interaction between the periplasmic substrate-binding protein (SBP) ModA and the transporter ModBC is dependent upon binding of molybdate. By contrast, vitamin B12 disrupts interactions between the transporter BtuCD and the SBP BtuF. Moreover, while ATP binds cooperatively to BtuCD-F, and acts synergistically with vitamin B12 to destabilize the BtuCD-F complex, no effect is observed for ATP binding on the stability of ModBC-A. These observations not only highlight the ability of mass spectrometry to capture these importer-SBP complexes but allow us to add molecular detail to proposed transport mechanisms.

Citations

Jul 15, 2020·Journal of Mass Spectrometry : JMS·Marie Barth, Carla Schmidt
Jul 9, 2020·Proceedings of the National Academy of Sciences of the United States of America·Jani Reddy BollaCarol V Robinson
Sep 10, 2020·Frontiers in Pharmacology·Francesco FiorentinoDante Rotili
May 7, 2020·Pflügers Archiv : European journal of physiology·Jean-Marc Jeckelmann, Bernhard Erni
Oct 25, 2020·FEBS Letters·Oded LewinsonMarkus A Seeger
Aug 24, 2019·Research in Microbiology·I Barry Holland
Nov 5, 2019·Research in Microbiology·Philippe Delepelaire
Nov 18, 2020·Nature Chemical Biology·Francesco FiorentinoCarol V Robinson
May 4, 2020·Journal of Proteomics·Elisabetta Boeri ErbaCarlo Petosa
Jul 17, 2021·Frontiers in Molecular Biosciences·James S DaviesRenwick C J Dobson
Sep 30, 2021·Chemical Communications : Chem Comm·Francesco FiorentinoCarol V Robinson

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Methods Mentioned

BETA
surface plasmon resonance
X-ray
microscale thermophoresis
fluorescence imaging
size exclusion chromatography

Software Mentioned

pymol
UniDec
PyMol Schrödinger LLC
Excel

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