TY  - JOUR
A1  - Vitt, Stella
A1  - Prinz, Simone
A1  - Hellwig, Nils
A1  - Morgner, Nina
A1  - Ermler, Ulrich
A1  - Buckel, Wolfgang
T1  - Molecular and low-resolution structural characterization of the Na+-translocating glutaconyl-CoA decarboxylase from clostridium symbiosum
T2  - Frontiers in microbiology
N2  - Some anaerobic bacteria use biotin-dependent Na+-translocating decarboxylases (Bdc) of β-keto acids or their thioester analogs as key enzymes in their energy metabolism. Glutaconyl-CoA decarboxylase (Gcd), a member of this protein family, drives the endergonic translocation of Na+ across the membrane with the exergonic decarboxylation of glutaconyl-CoA (ΔG0’ ≈−30 kJ/mol) to crotonyl-CoA. Here, we report on the molecular characterization of Gcd from Clostridium symbiosum based on native PAGE, size exclusion chromatography (SEC) and laser-induced liquid bead ion desorption mass spectrometry (LILBID-MS). The obtained molecular mass of ca. 400 kDa fits to the DNA sequence-derived mass of 379 kDa with a subunit composition of 4 GcdA (65 kDa), 2 GcdB (35 kDa), GcdC1 (15 kDa), GcdC2 (14 kDa), and 2 GcdD (10 kDa). Low-resolution structural information was achieved from preliminary electron microscopic (EM) measurements, which resulted in a 3D reconstruction model based on negative-stained particles. The Gcd structure is built up of a membrane-spanning base primarily composed of the GcdB dimer and a solvent-exposed head with the GcdA tetramer as major component. Both globular parts are bridged by a linker presumably built up of segments of GcdC1, GcdC2 and the 2 GcdDs. The structure of the highly mobile Gcd complex represents a template for the global architecture of the Bdc family.
KW  - anaerobic energy metabolism
KW  - glutaconyl-CoA decarboxylase
KW  - ion translocation
KW  - biotin
KW  - negative-stain electron microscopy
KW  - LILBID-MS
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/53350
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-533505
N1  - Copyright © 2020 Vitt, Prinz, Hellwig, Morgner, Ermler and Buckel. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
VL  - 11
IS  - 480
PB  - Frontiers Research Foundation
CY  - Lausanne
ER  -