Structure of the bongkrekic acid-inhibited mitochondrial ADP/ATP carrier

DIPHOSPHATIDYL GLYCEROL; BIS-(1,2-DIACYL-SN-GLYCERO-3-PHOSPHO)-1',3'-SN-GLYCEROL

The Molecular Mechanism of Transport by the Mitochondrial ADP/ATP Carrier.

Nanobody selected against the BKA-inhibited ADP/ATP carrier

MSNKQETKILGMPPFVVDFLMGGVSAAVSKTAAAPIERIKLLVQNQDEMIKAGRLDRRYNGIIDCFRRTTADEGLMALWR
GNTANVIRYFPTQALNFAFRDKFKAMFGYKKDKDGYAKWMAGNLASGGAAGATSLLFVYSLDYARTRLANDAKSAKGGGA
RQFNGLIDVYRKTLASDGIAGLYRGFGPSVAGIVVYRGLYFGMYDSIKPVVLVGPLANNFLASFLLGWCVTTGAGIASYP
LDTVRRRMMMTSGEAVKYKSSIDAFRQIIAKEGVKSLFKGAGANILRGVAGAGVLSIYDQLKILLFGKAFKGGSG

QVQLVESGGGLVQAGDSLRLACAASGITFSSYTMGWFRQAPGKEREYIARITGSGSNTYYADSVKGRYTISRDNAKNTAY
LQMNSLKPEDTANYYCAARDDGRMYQGFYDFEYWGQGTQVTVSSDRAIEGR

Thermothelomyces thermophila (strain ATCC 42464 / BCRC 31852 / DSM 1799)

0.1M MES pH 6.5, 1% 3-methyl-3-pentanol, 22% PEG400

Medical Research Council (United Kingdom)

(A) Crystal diffracted anisotropically, and data was treated with STARANISO.
(B) Fourier map coefficients have been deposited along with the observed data.
(C) A PEG molecule lies on a symmetry axis (special position) and has been modelled with occupancy=0.5.
(D) Some cardiolipins molecules have been modelled with partial occupancy, as described in the accompanying paper.
(E) Met250-Ser252 and Lys257 lie in very weak electron density. Their position is supported by good electron density obtained from an earlier crystal.
(F) There are unmodelled bits of electron density near the N-terminus of the carrier, which are likely to be lipid headgroups/tails.
(G) Additional electron density at the C-terminus of the carrier indicates where the C-terminal tail is likely to run, but it is not possible to model this accurately.
(H) The structure factor file contains a second data block containing the structure factors from a crystal of BKA-inhibited TtAac grown without the nanobody. This P212121 crystal diffracted to lower resolution and with significant anisotropy, but was used to confirm the domain positions observed in the P3221 crystal of the TtAac-Nb complex. Full details can be found in the primary citation.

Transporter, Inhibitor, Mitochondrial, Carrier, MEMBRANE PROTEIN

MSNKQETKILGMPPFVVDFLMGGVSAAVSKTAAAPIERIKLLVQNQDEMIKAGRLDRRYNGIIDCFRRTTADEGLMALWR
GNTANVIRYFPTQALNFAFRDKFKAMFGYKKDKDGYAKWMAGNLASGGAAGATSLLFVYSLDYARTRLANDAKSAKGGGA
RQFNGLIDVYRKTLASDGIAGLYRGFGPSVAGIVVYRGLYFGMYDSIKPVVLVGPLANNFLASFLLGWCVTTGAGIASYP
LDTVRRRMMMTSGEAVKYKSSIDAFRQIIAKEGVKSLFKGAGANILRGVAGAGVLSIYDQLQILLFGKAFKGGSG