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TitleNative mass spectrometry and structural studies reveal modulation of MsbA-nucleotide interactions by lipids.
Journal, issue, pagesNat Commun, Vol. 15, Issue 1, Page 5946, Year 2024
Publish dateJul 15, 2024
AuthorsTianqi Zhang / Jixing Lyu / Bowei Yang / Sangho D Yun / Elena Scott / Minglei Zhao / Arthur Laganowsky /
PubMed AbstractThe ATP-binding cassette (ABC) transporter, MsbA, plays a pivotal role in lipopolysaccharide (LPS) biogenesis by facilitating the transport of the LPS precursor lipooligosaccharide (LOS) from the ...The ATP-binding cassette (ABC) transporter, MsbA, plays a pivotal role in lipopolysaccharide (LPS) biogenesis by facilitating the transport of the LPS precursor lipooligosaccharide (LOS) from the cytoplasmic to the periplasmic leaflet of the inner membrane. Despite multiple studies shedding light on MsbA, the role of lipids in modulating MsbA-nucleotide interactions remains poorly understood. Here we use native mass spectrometry (MS) to investigate and resolve nucleotide and lipid binding to MsbA, demonstrating that the transporter has a higher affinity for adenosine 5'-diphosphate (ADP). Moreover, native MS shows the LPS-precursor 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo)-lipid A (KDL) can tune the selectivity of MsbA for adenosine 5'-triphosphate (ATP) over ADP. Guided by these studies, four open, inward-facing structures of MsbA are determined that vary in their openness. We also report a 2.7 Å-resolution structure of MsbA in an open, outward-facing conformation that is not only bound to KDL at the exterior site, but with the nucleotide binding domains (NBDs) adopting a distinct nucleotide-free structure. The results obtained from this study offer valuable insight and snapshots of MsbA during the transport cycle.
External linksNat Commun / PubMed:39009687 / PubMed Central
MethodsEM (single particle)
Resolution2.68 - 3.9 Å
Structure data

41596

8tso

EMDB-41596, PDB-8tso:
KDL bound, nucleotide-free MsbA in open, outward-facing conformation
Method: EM (single particle) / Resolution: 2.68 Å

41597

8tsp

EMDB-41597, PDB-8tsp:
Open, inward-facing MsbA structure (OIF1)
Method: EM (single particle) / Resolution: 3.9 Å

41598

8tsq

EMDB-41598, PDB-8tsq:
Open, inward-facing MsbA structure (OIF2)
Method: EM (single particle) / Resolution: 3.6 Å

41599

8tsr

EMDB-41599, PDB-8tsr:
Open, inward-facing MsbA structure (OIF4)
Method: EM (single particle) / Resolution: 3.9 Å

41600

8tss

EMDB-41600, PDB-8tss:
Open, inward-facing MsbA structure (OIF3)
Method: EM (single particle) / Resolution: 3.7 Å

Chemicals

ChemComp-CXE:
PENTAETHYLENE GLYCOL MONODECYL ETHER

ChemComp-KDL:
(2~{R},4~{R},5~{R},6~{R})-6-[(1~{R})-1,2-bis(oxidanyl)ethyl]-2-[(2~{R},4~{R},5~{R},6~{R})-6-[(1~{R})-1,2-bis(oxidanyl)ethyl]-2-carboxy-2-[[(2~{R},3~{S},4~{R},5~{R},6~{R})-5-[[(3~{R})-3-dodecanoyloxytetradecanoyl]amino]-6-[[(2~{R},3~{S},4~{R},5~{R},6~{R})-3-oxidanyl-5-[[(3~{R})-3-oxidanyltetradecanoyl]amino]-4-[(3~{R})-3-oxidanyltetradecanoyl]oxy-6-phosphonooxy-oxan-2-yl]methoxy]-3-phosphonooxy-4-[(3~{R})-3-tetradecanoyloxytetradecanoyl]oxy-oxan-2-yl]methoxy]-5-oxidanyl-oxan-4-yl]oxy-4,5-bis(oxidanyl)oxane-2-carboxylic acid

Source
  • escherichia coli (E. coli)
KeywordsTRANSPORT PROTEIN / ABC transporter

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